Found 294 structures.
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1. Compound ID: 107
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136105,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_225177,IEDB_885813,IEDB_885823,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 20
Bousquet E, Khitri M, Lay L, Nicotra F, Panza L, Russo G "Capsular polysaccharide of Streptococcus pneumoniae type 19F: synthesis of the repeating unit" -
Carbohydrate Research 311(4) (1998) 171-181
A new and more versatile synthesis of β-D-ManpNAc-(1→4)-α-D-Glcp-(1→2)-α-L-Rhap, the trisaccharide repeating unit of the Streptococcus pneumoniae type 19F capsular polysaccharide, is described. The present approach allows a simple access to different fragments containing the trisaccharide and the conjugation of the product(s) to a protein through the selective manipulation of the anomeric position at the reducing end and of the HO-4 function at the nonreducing end. The synthetic scheme shows an efficient application of the sulfoxide method for the stereoselective and high yielding formation of the glycosidic linkages
synthesis, polysaccharide, Streptococcus, Streptococcus pneumoniae, repeating unit, Oligosaccharides, Anomeric sulfoxides, glycosylationcapsular, capsular polysaccharide, glycoconjugate vaccines, sulfoxides, type
NCBI PubMed ID: 9825520Publication DOI: 10.1016/s0008-62159800218-3Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Dipartimento di Chimica Organica e Industriale, via Venezian 21, I-20133, Milano, Italy
Methods: 13C NMR, 1H NMR, TLC
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2. Compound ID: 405
|
R-3HOBut-(1-6)-+ a-L-Fucp-(1-2)-+
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-3)-a-D-FucpNAc4N-(1-2)-a-D-Hepp-(1-3)-b-D-ManpNAc-(1-4)-b-D-Quip3NAc-(1- |
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Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136045,IEDB_142345,IEDB_142489,IEDB_144562,IEDB_152214,IEDB_174333,IEDB_885813,SB_86
The structure is contained in the following publication(s):
- Article ID: 135
Tzianabos A, Wang JY, Kasper DL "Biological chemistry of immunomodulation by zwitterionic polysaccharides" -
Carbohydrate Research 338(23) (2003) 2531-2538
Capsular polysaccharides isolated from pathogenic bacteria are comprised typically of many repeating units from one to eight or more monosaccharides in length. These polysaccharides stimulate the murine humoral immune system to elicit primarily IgM antibody responses. Studies conducted primarily in the mouse have characterized these polymers as T cell-independent antigens. These mouse studies and the relatively poor immunogenicity of polysaccharides in human hosts have led to the design of vaccines by coupling these polysaccharides to protein carriers to stimulate a T cell-dependent response. However, a newly described class of bacterial polysaccharides has been characterized that have the ability to modulate the cellular immune system. They are structurally diverse, but all share a zwitterionic charge motif that allows them to directly interact with T cells and antigen-presenting cells to initiate an immunomodulatory T cell response. These polymers, termed zwitterionic polysaccharides (ZPSs), elicit T cell-derived chemokines and cytokines that influence the immune response governing at least one classic host response to bacterial infection: abscess formation. This review will describe the biological and structural aspects of ZPSs that convey these activities.
T cell, polysaccharides, structure/function
NCBI PubMed ID: 14670714Publication DOI: 10.1016/j.carres.2003.06.005Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: atzianabos@channing.harvard.edu
Institutions: Department of Medicine, Channing Laboratory, 181 Longwood Ave., Brigham and Women's Hospital, Boston, MA 02115, USA
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3. Compound ID: 418
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-4)-a-D-GlcpA2Ac3Ac-(1-3)-a-D-Galp-(1-3)-b-D-ManpNAc4Ac6Ac-(1-4)-b-D-Glcp-(1-4)-a-D-Glcp2Ac3Ac-(1- |
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Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_115136,IEDB_136906,IEDB_137472,IEDB_140630,IEDB_141794,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_885813,IEDB_983931,SB_192,SB_61,SB_7
The structure is contained in the following publication(s):
- Article ID: 147
van Selm S, Kolkman MA, van der Zeijst BA, Zwaagstra KA, Gaastra W, van Putten JP "Organization and characterization of the capsule biosynthesis locus of Streptococcus pneumoniae serotype 9V" -
Microbiology 148(6) (2002) 1747-1755
The capsular polysaccharide (CPS) synthesis locus of Streptococcus pneumoniae serotype 9V was amplified by long-range PCR and sequenced. The locus was 17368 bp in size and contained 15 ORFs. The genetic organization of the cluster shared many features with other S. pneumoniae capsule loci, including the presence of four putative regulatory genes at the 5' end. Comparative sequence analyses allowed putative functions to be assigned to each of the gene products. The ORFs appeared to encode, besides the four regulatory genes, five glycosyltransferases, two O-acetyltransferases, an N-acetylglucosamine 2-epimerase, a glucose 6-dehydrogenase, an oligosaccharide transporter protein and a polysaccharide repeating unit polymerase. These functions covered the steps proposed in the CPS biosynthesis of serotype 9V. TLC of carbohydrate intermediates formed after incubation of bacterial membrane preparations with 14C-labelled precursors demonstrated that the fifth ORF (cps9vE) encoded a UDP-glucosyl-1-phosphate transferase. This function was confirmed with the help of a cps9vE mutant that carried a deletion of a guanine residue located adjacent to a stretch of adenines. The identification and characterization of the serotype 9V locus is a major step in unravelling the 9V capsule biosynthesis pathway and broadens the insight into the genetic diversity of the S. pneumoniae capsule loci
capsular polysaccharide biosynthesis, glucosyltransferase
NCBI PubMed ID: 12055294Journal NLM ID: 0376646Publisher: Washington, DC: Kluwer Academic/Plenum Publishers
Correspondence: j.vanputten@vet.uu.nl
Institutions: Bacteriology Division, Department of Infectious Diseases and Immunology, Utrecht University, PO Box 80.165, 3508 TD Utrecht, The Netherlands, Genencor International B. V., 2300 AE Leiden, The Netherlands, National Institute of Public Health and the Environment, 3720 BA Bilthoven, The Netherlands
- Article ID: 294
Kolberg J, Jones C "Monoclonal antibodies with specificities for Streptococcus pneumoniae group 9 capsular polysaccharides" -
FEMS Immunology and Medical Microbiology 20(4) (1998) 249-255
Streptococcus pneumoniae group 9 includes four capsular polysaccharide types: 9A, 9L, 9N and 9V. We have generated four mouse monoclonal antibodies against group 9 polysaccharide using heat-treated S. pneumoniae strains of different capsular polysaccharides types as immunogens. The specificities of the monoclonal antibodies were determined by ELISA using capsular polysaccharide directly coated to the wells as antigens and by dot blotting with heat-treated bacteria. Two groups of monoclonal antibodies were found. The first group included two monoclonal antibodies which were found to be capsular type specific. The second group was monoclonal antibodies that bound to epitopes shared by two or three pneumococcal group 9 types. The monoclonal antibody 204,A-4 (IgM) was found to be specific for S. pneumoniae type 9N. The binding of the type 9V specific monoclonal antibody 206,F-5 (IgG1) was found to be dependent upon O-acetyl groups. Monoclonal antibody 205,F-3 (IgM) reacted also with type 9V, but was found to cross-react with types 9A and 9L. The binding of this monoclonal antibody to polysaccharide 9V was not dependent upon O-acetyl moieties. The fourth monoclonal antibody (214,G-5, isotype IgM) did not show any correlation between reactivity with isolated polysaccharides and dot blotting with relevant bacteria. The monoclonal antibody reacted with polysaccharides 9A and 9L in ELISA, but not with the homologous bacteria.
Streptococcus pneumoniae, capsular polysaccharides, monoclonal antibodies, group 9 polysaccharide
NCBI PubMed ID: 9626929Journal NLM ID: 9315554Publisher: Elsevier
Institutions: National Institute of Public Health, Department of Vaccinology, P.O. Box 4404 Torshov, N-0403 Oslo, Norway, National Institute for Biological Standards and Control, South Mimms, Herts. EN6 3QG, UK.
- Article ID: 4284
Calix JJ, Saad JS, Brady AM, Nahm MH "Structural characterization of Streptococcus pneumoniae serotype 9A capsule polysaccharide reveals role of glycosyl 6-O-acetyltransferase wcjE in serotype 9V capsule biosynthesis and immunogenicity" -
Journal of Biological Chemistry 287(17) (2012) 13996-14003
The putative capsule O-acetyltransferase gene wcjE is highly conserved across various Streptococcus pneumoniae serotypes, but the role of the gene in capsule biosynthesis and bacterial fitness remains largely unclear. Isolates expressing pneumococcal serotype 9A arise from precursors expressing wcjE-associated serotype 9V through loss-of-function mutation to wcjE. To define the biosynthetic role of 9V wcjE, we characterized the structure and serological properties of serotype 9V and 9A capsule polysaccharide (PS). NMR data revealed that both 9V and 9A PS are composed of an identical pentasaccharide repeat unit, as reported previously. However, in sharp contrast to previous studies on 9A PS being devoid of any O-acetylation, we identified O-acetylation of α-glucuronic acid and α-glucose in 9A PS. In addition, 9V PS also contained -CH(2) O-acetylation of β-N-acetylmannosamine, a modification that disappeared following in vitro recombinatorial deletion of wcjE. We also show that serotyping sera and monoclonal antibodies specific for 9V and 9A bound capsule PS in an O-acetate-dependent manner. Furthermore, IgG and to a lesser extent IgM from human donors immunized with serotype 9V PS displayed stronger binding to 9V compared with 9A PS. We conclude that serotype 9V wcjE mediates 6-O-acetylation of β-N-acetylmannosamine. This PS modification can be selectively targeted by antibodies in immunized individuals, identifying a potential selective advantage for wcjE inactivation and serotype 9A emergence.
structure, capsule polysaccharide, capsule biosynthesis, O-acetyltransferase, Streptococcus pneumoniae 9A, wcjE
NCBI PubMed ID: 22367197Publication DOI: 10.1074/jbc.M112.346924Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: nahm@uab.edu
Institutions: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA
Methods: 13C NMR, 1H NMR, NMR-2D, GC-MS, sugar analysis, ELISA, de-O-acetylation, NMR-1D, serological methods
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4. Compound ID: 698
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b-L-Arap4N-(1--P--6)--a-D-GlcpN-(1-4)-a-Kdop-(2-4)-+
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b-D-GlcpNAc-(1-4)-+ a-D-6dTalp2Ac3Me-(1-3)-+ a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Galp-(1-2)-+ |
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a-D-GlcpNAc-(1-3)-a-D-Fucp-(1-2)-b-D-GlcpA-(1-4)-a-D-Fucp-(1-3)-a-D-GalpNAc-(1-4)-b-D-ManpNAc-(1-4)-a-D-Galp-(1-3)-b-D-GalpNAc-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-6)-a-D-Manp-(1-4)-a-D-Manp-(1-5)-a-Kdop-(2-6)-b-D-GlcpN-(1-6)-a-D-GlcpN-(1-P
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a-Kdop-(2-3)-+ |
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Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_115015,IEDB_115136,IEDB_130648,IEDB_130650,IEDB_130659,IEDB_130701,IEDB_135813,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140116,IEDB_140630,IEDB_141584,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144983,IEDB_144990,IEDB_144998,IEDB_146664,IEDB_149135,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_167071,IEDB_190606,IEDB_232584,IEDB_423153,IEDB_885813,IEDB_885822,IEDB_983930,IEDB_983931,SB_192,SB_198,SB_44,SB_67,SB_7,SB_72,SB_86
The structure is contained in the following publication(s):
- Article ID: 187
Vinogradov E, Petersen BO, Sadovskaya I, Jabbouri S, Duus J, Helander IM "Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E-82164" -
European Journal of Biochemistry 270(14) (2003) 3036-3046
The structures of the oligosaccharides obtained after acetic acid hydrolysis and alkaline deacylation of the rough-type lipopolysaccharide (LPS) from Pectinatus frisingensis strain VTT E-82164 were analysed using NMR spectroscopy, MS and chemical methods. The LPS contains two major structural variants, differing by a decasaccharide fragment, and some minor variants lacking the terminal glucose residue. The largest structure of the carbohydrate backbone of the LPS that could be deduced from experimental results consists of 25 monosaccharides (including the previously found Arap4NP residue in lipid A) arranged in a well-defined nonrepetitive structure: We presume that the shorter variant with R1 = H represents the core-lipid A part of the LPS, and the additional fragment is present instead of the O-specific polysaccharide. Structures of this type have not been previously described. Analysis of the deacylation products obtained from the LPS of the smooth strain, VTT E-79100T, showed that it contains a very similar core but with one different glycosidic linkage.
Lipopolysaccharide, structure, core, strain, carbohydrate, lipid A, backbone, Pectinatus
NCBI PubMed ID: 12846837Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: evguenii.vinogradov@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark, Laboratoire de Recherche sur les Biomateriaux et Biotechnologies, Universite de Littoral-Cote d'Opale, Bassin Napoleon BP 120, Boulogne-sur-mer, France, Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Finland
Methods: NMR-2D, NMR, chemical methods, MS
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5. Compound ID: 701
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a-D-GlcpN-(1-4)-a-Kdop-(2-4)-+
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b-D-GlcpN-(1-4)-+ a-D-6dTalp3Me-(1-3)-+ a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Galp-(1-2)-+ |
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a-D-GlcpN-(1-3)-a-D-Fucp-(1-2)-b-D-GlcpA-(1-4)-a-D-Fucp-(1-3)-a-D-GalpN-(1-4)-b-D-ManpN-(1-4)-a-D-Galp-(1-3)-b-D-GalpN-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-6)-a-D-Manp-(1-4)-a-D-Manp-(1-5)-a-Kdop-(2-6)-b-D-GlcpN-(1-6)-a-D-GlcpN-(1-P
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a-Kdop-(2-3)-+ |
Show graphically |
Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_115015,IEDB_115136,IEDB_130650,IEDB_130659,IEDB_130701,IEDB_136906,IEDB_137472,IEDB_137473,IEDB_140116,IEDB_140630,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144983,IEDB_144998,IEDB_146664,IEDB_149135,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_190606,IEDB_232584,IEDB_423153,IEDB_983930,IEDB_983931,SB_192,SB_198,SB_44,SB_67,SB_7,SB_72,SB_86
The structure is contained in the following publication(s):
- Article ID: 187
Vinogradov E, Petersen BO, Sadovskaya I, Jabbouri S, Duus J, Helander IM "Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E-82164" -
European Journal of Biochemistry 270(14) (2003) 3036-3046
The structures of the oligosaccharides obtained after acetic acid hydrolysis and alkaline deacylation of the rough-type lipopolysaccharide (LPS) from Pectinatus frisingensis strain VTT E-82164 were analysed using NMR spectroscopy, MS and chemical methods. The LPS contains two major structural variants, differing by a decasaccharide fragment, and some minor variants lacking the terminal glucose residue. The largest structure of the carbohydrate backbone of the LPS that could be deduced from experimental results consists of 25 monosaccharides (including the previously found Arap4NP residue in lipid A) arranged in a well-defined nonrepetitive structure: We presume that the shorter variant with R1 = H represents the core-lipid A part of the LPS, and the additional fragment is present instead of the O-specific polysaccharide. Structures of this type have not been previously described. Analysis of the deacylation products obtained from the LPS of the smooth strain, VTT E-79100T, showed that it contains a very similar core but with one different glycosidic linkage.
Lipopolysaccharide, structure, core, strain, carbohydrate, lipid A, backbone, Pectinatus
NCBI PubMed ID: 12846837Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: evguenii.vinogradov@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark, Laboratoire de Recherche sur les Biomateriaux et Biotechnologies, Universite de Littoral-Cote d'Opale, Bassin Napoleon BP 120, Boulogne-sur-mer, France, Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Finland
Methods: NMR-2D, NMR, chemical methods, MS
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6. Compound ID: 702
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a-D-GlcpN-(1-4)-a-Kdop-(2-4)-+
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b-D-GlcpN-(1-4)-+ a-D-6dTalp3Me-(1-3)-+ a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Galp-(1-2)-+ |
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a-D-GlcpN-(1-3)-a-D-Fucp-(1-2)-a-D-GlcpA-(1-4)-a-D-Fucp-(1-3)-a-D-GalpN-(1-4)-b-D-ManpN-(1-4)-a-D-Galp-(1-3)-b-D-GalpN-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-6)-a-D-Manp-(1-4)-a-D-Manp-(1-5)-a-Kdop-(2-6)-b-D-GlcpN-(1-6)-a-D-GlcpN-(1-P
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a-Kdop-(2-3)-+ |
Show graphically |
Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_115015,IEDB_115136,IEDB_130650,IEDB_130659,IEDB_130701,IEDB_136906,IEDB_137472,IEDB_137473,IEDB_140116,IEDB_140630,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144983,IEDB_144998,IEDB_146664,IEDB_149135,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_190606,IEDB_232584,IEDB_983930,IEDB_983931,SB_192,SB_198,SB_44,SB_67,SB_7,SB_72,SB_86
The structure is contained in the following publication(s):
- Article ID: 187
Vinogradov E, Petersen BO, Sadovskaya I, Jabbouri S, Duus J, Helander IM "Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E-82164" -
European Journal of Biochemistry 270(14) (2003) 3036-3046
The structures of the oligosaccharides obtained after acetic acid hydrolysis and alkaline deacylation of the rough-type lipopolysaccharide (LPS) from Pectinatus frisingensis strain VTT E-82164 were analysed using NMR spectroscopy, MS and chemical methods. The LPS contains two major structural variants, differing by a decasaccharide fragment, and some minor variants lacking the terminal glucose residue. The largest structure of the carbohydrate backbone of the LPS that could be deduced from experimental results consists of 25 monosaccharides (including the previously found Arap4NP residue in lipid A) arranged in a well-defined nonrepetitive structure: We presume that the shorter variant with R1 = H represents the core-lipid A part of the LPS, and the additional fragment is present instead of the O-specific polysaccharide. Structures of this type have not been previously described. Analysis of the deacylation products obtained from the LPS of the smooth strain, VTT E-79100T, showed that it contains a very similar core but with one different glycosidic linkage.
Lipopolysaccharide, structure, core, strain, carbohydrate, lipid A, backbone, Pectinatus
NCBI PubMed ID: 12846837Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: evguenii.vinogradov@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark, Laboratoire de Recherche sur les Biomateriaux et Biotechnologies, Universite de Littoral-Cote d'Opale, Bassin Napoleon BP 120, Boulogne-sur-mer, France, Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Finland
Methods: NMR-2D, NMR, chemical methods, MS
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7. Compound ID: 706
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b-D-GlcpNAc-(1-4)-+ a-D-6dTalp3Me-(1-3)-+ a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Galp-(1-2)-+
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a-D-GlcpNAc-(1-3)-a-D-Fucp-(1-2)-b-D-GlcpA-(1-4)-a-D-Fucp-(1-3)-a-D-GalpNAc-(1-4)-b-D-ManpNAc-(1-4)-a-D-Galp-(1-3)-b-D-GalpNAc-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-6)-a-D-Manp-(1-4)-a-D-Manp-(1-5)-Kdo-ol |
Show graphically |
Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_115015,IEDB_115136,IEDB_130648,IEDB_130701,IEDB_135813,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140116,IEDB_140630,IEDB_141584,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144983,IEDB_144990,IEDB_144998,IEDB_146664,IEDB_149135,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_167071,IEDB_190606,IEDB_232584,IEDB_423153,IEDB_885813,IEDB_885822,IEDB_983930,IEDB_983931,SB_192,SB_198,SB_44,SB_67,SB_7,SB_72,SB_86
The structure is contained in the following publication(s):
- Article ID: 187
Vinogradov E, Petersen BO, Sadovskaya I, Jabbouri S, Duus J, Helander IM "Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E-82164" -
European Journal of Biochemistry 270(14) (2003) 3036-3046
The structures of the oligosaccharides obtained after acetic acid hydrolysis and alkaline deacylation of the rough-type lipopolysaccharide (LPS) from Pectinatus frisingensis strain VTT E-82164 were analysed using NMR spectroscopy, MS and chemical methods. The LPS contains two major structural variants, differing by a decasaccharide fragment, and some minor variants lacking the terminal glucose residue. The largest structure of the carbohydrate backbone of the LPS that could be deduced from experimental results consists of 25 monosaccharides (including the previously found Arap4NP residue in lipid A) arranged in a well-defined nonrepetitive structure: We presume that the shorter variant with R1 = H represents the core-lipid A part of the LPS, and the additional fragment is present instead of the O-specific polysaccharide. Structures of this type have not been previously described. Analysis of the deacylation products obtained from the LPS of the smooth strain, VTT E-79100T, showed that it contains a very similar core but with one different glycosidic linkage.
Lipopolysaccharide, structure, core, strain, carbohydrate, lipid A, backbone, Pectinatus
NCBI PubMed ID: 12846837Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: evguenii.vinogradov@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark, Laboratoire de Recherche sur les Biomateriaux et Biotechnologies, Universite de Littoral-Cote d'Opale, Bassin Napoleon BP 120, Boulogne-sur-mer, France, Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Finland
Methods: NMR-2D, NMR, chemical methods, MS
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8. Compound ID: 707
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b-D-GlcpNAc-(1-4)-+ a-D-6dTalp2Ac3Me-(1-3)-+ a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Galp-(1-2)-+
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a-D-GlcpNAc-(1-3)-a-D-Fucp-(1-2)-b-D-GlcpA-(1-4)-a-D-Fucp-(1-3)-a-D-GalpNAc-(1-4)-b-D-ManpNAc-(1-4)-a-D-Galp-(1-3)-b-D-GalpNAc-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-6)-a-D-Manp-(1-4)-a-D-Manp-(1-5)-Kdo |
Show graphically |
Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_115015,IEDB_115136,IEDB_130648,IEDB_130650,IEDB_130701,IEDB_135813,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140116,IEDB_140630,IEDB_141584,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144983,IEDB_144990,IEDB_144998,IEDB_146664,IEDB_149135,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_167071,IEDB_190606,IEDB_232584,IEDB_423153,IEDB_885813,IEDB_885822,IEDB_983930,IEDB_983931,SB_192,SB_198,SB_44,SB_67,SB_7,SB_72,SB_86
The structure is contained in the following publication(s):
- Article ID: 187
Vinogradov E, Petersen BO, Sadovskaya I, Jabbouri S, Duus J, Helander IM "Structure of the exceptionally large nonrepetitive carbohydrate backbone of the lipopolysaccharide of Pectinatus frisingensis strain VTT E-82164" -
European Journal of Biochemistry 270(14) (2003) 3036-3046
The structures of the oligosaccharides obtained after acetic acid hydrolysis and alkaline deacylation of the rough-type lipopolysaccharide (LPS) from Pectinatus frisingensis strain VTT E-82164 were analysed using NMR spectroscopy, MS and chemical methods. The LPS contains two major structural variants, differing by a decasaccharide fragment, and some minor variants lacking the terminal glucose residue. The largest structure of the carbohydrate backbone of the LPS that could be deduced from experimental results consists of 25 monosaccharides (including the previously found Arap4NP residue in lipid A) arranged in a well-defined nonrepetitive structure: We presume that the shorter variant with R1 = H represents the core-lipid A part of the LPS, and the additional fragment is present instead of the O-specific polysaccharide. Structures of this type have not been previously described. Analysis of the deacylation products obtained from the LPS of the smooth strain, VTT E-79100T, showed that it contains a very similar core but with one different glycosidic linkage.
Lipopolysaccharide, structure, core, strain, carbohydrate, lipid A, backbone, Pectinatus
NCBI PubMed ID: 12846837Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: evguenii.vinogradov@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark, Laboratoire de Recherche sur les Biomateriaux et Biotechnologies, Universite de Littoral-Cote d'Opale, Bassin Napoleon BP 120, Boulogne-sur-mer, France, Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Finland
Methods: NMR-2D, NMR, chemical methods, MS
Expand this compound
Collapse this compound
9. Compound ID: 815
Structure type: polymer chemical repeating unit
Trivial name: poly(glycosyl phosphate)
Compound class: CPS
Contained glycoepitopes: IEDB_136105,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_158557,IEDB_225177,IEDB_885813,IEDB_885823,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 219
Whitfield C, Paiment A "Biosynthesis and assembly of group 1 capsular polysaccharides in Escherichia coli and related extracellular polysaccharides in other bacteria" -
Carbohydrate Research 338(23) (2003) 2491-2502
Extracellular and capsular polysaccharides (EPSs and CPSs) are produced by a wide range of bacteria, including important pathogens of humans, livestock, and plants. These polymers are major surface antigens and serve a variety of roles in virulence, depending on the biology of the producing organism. In addition to their importance in disease, some EPSs also have industrial applications as gelling and emulsifying agents. An understanding of the processes involved in the synthesis and regulation of CPSs and EPSs therefore potentially contributes to an understanding of the disease state, surface expression of protective antigens, and modulation of polymer structure to give defined physical properties. Escherichia coli has provided important model systems for EPS and CPS biosynthesis. Here we describe current knowledge concerning assembly of the Group 1 CPSs of E. coli and the conservation of similar mechanisms in other bacteria
biosynthesis, Escherichia coli, capsular polysaccharide, extracellular polysaccharide, K-antigens, Wzy-dependent assembly
Publication DOI: 10.1016/j.carres.2003.08.010Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: cwhitfie@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
- Article ID: 279
Kaji E, Osa Y, Tanaike M, Hosokawa Y, Takayanagi H, Takada A "An alternative access to a trisaccharide repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 19A" -
Chemical and Pharmaceutical Bulletin 44 (1996) 437-440
A chemical synthesis has been achieved for β-D-ManNAc-(1→4)-α-D-Glc-(1→3)-L-Rha, a trisaccharide repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 19A, by stepwise link-up of the suitably functionalized, constituent sugar units. A beta-selective glycosylation of trimethylsilylethyl glucoside having free 4-OH with 2-(benzoyloxyimino)-2-deoxyglycosyl bromide, followed by manno-selective hydroboration, N-acetylation, and functionalization of the anomeric center (1-OSE→1-OH→1-F), gave a key disaccharide donor, β-D-ManNAc-(1→4)-α-D-Glc-(1→F. Ensuing glycosylation of an L-rhamnosyl acceptor with the donor substrate afforded, after deblocking, the target trisaccharide in 6.5% yield over 13 steps from D-glucose.
capsular polysaccharide, glycosylation, Streptococcus pneumoniae type 19A, 2-ulose oxime, β-D-mannosaminide, hydroboration
NCBI PubMed ID: 8998845Journal NLM ID: 0377775Publisher: Pharmaceutical Society Of Japan
Institutions: School of Pharmaceutical Sciences, Kitasato University, Japan, School of Pharmaceutical Sciences, Kitasato University
Methods: 13C NMR, 1H NMR, FAB-MS, TLC, chemical synthesis, chemical methods, UV, glycosylation
- Article ID: 332
Morona JK, Morona R, Paton JC "Comparative genetics of capsular polysaccharide biosynthesis in Streptococcus pneumoniae types belonging to serogroup 19" -
Journal of Bacteriology 181(17) (1999) 5355-5364
The genetic basis for the structural diversity of capsule polysaccharide (CPS) in Streptococcus pneumoniae serogroup 19 (consisting of types 19F, 19A, 19B, and 19C) has been determined for the first time. In this study, the genetic basis for the 19A and 19C serotypes is described, and the structures of all four serogroup 19 cps loci and their flanking sequences are compared. Transformation studies show that the structural difference between the 19A and 19F CPSs is likely to be a consequence of differences between their respective polysaccharide polymerase genes (cps19aI and cps19fI). The CPS of type 19C differs from that of type 19B by the addition of glucose. We have identified a single gene difference between the two cps loci (cps19cS), which is likely to encode a glucosyl transferase. The arrangement of the genes within the cps19 loci is highly conserved, with 13 genes (cps19A to -H and cps19K to -O) common to all four serogroup 19 members. These cps genes encode functions required for the synthesis of the shared trisaccharide component of the group 19 CPS repeat unit structures. Furthermore, the genetic differences between the group 19 cps loci identified are consistent with the CPS structures of the individual serotypes. Functions have been assigned to nearly all of the cps19 gene products, based on either gene complementation or similarity to other proteins with known functions, and putative biosynthetic pathways for production of all four group 19 CPSs have been proposed.
biosynthesis, genetic, Streptococcus, Streptococcus pneumoniae, capsular polysaccharide, type, serogroup
NCBI PubMed ID: 10464207Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: patonj@wch.sa.gov.au
Institutions: Molecular Microbiology Unit, Women's and Children's Hospital, North Adelaide, South Australia 5006, Department of Microbiology and Immunology, University of Adelaide, Adelaide, South Australia 5005
Methods: PCR, DNA sequencing, Southern blotting
- Article ID: 377
Sheng SQ, Cherniak R "Identification of phosphorylation sites in polysaccharides by 1- D 1H-31P HMQC experiments" -
Journal of Carbohydrate Chemistry 17(2) (1998) 317-321
polysaccharide, identification, site, phosphorylation, HMQC
Publication DOI: 10.1080/07328309808002331Journal NLM ID: 8218151Publisher: Marcel Dekker
Institutions: Department of Chemistry (LBCS), Georgia State University, Atlanta, GA 30303-3083, USA
Methods: 1H NMR, NMR-2D, 31P NMR
- Article ID: 507
Pujar NS, Huang NF, Daniels CL, Dieter L, Gayton MG, Lee AL "Base hydrolysis of phosphodiester bonds in pneumococcal polysaccharides" -
Biopolymers 75(1) (2004) 71-84
A comprehensive study of the base hydrolysis of all phosphodiester bond-containing capsular polysaccharides of the 23-valent pneumococcal vaccine is described here. Capsular polysaccharides from serotypes 6B, 10A, 17F, 19A, 19F, and 20 contain a phosphodiester bond that connects the repeating units in these polysaccharides (also referred to as backbone phosphodiester bonds), and polysaccharides from serotypes 11A, 15B, 18C, and 23F contain a phosphodiester bond that links a side chain to their repeating units. Molecular weight measurements of the polysaccharides, using high performance size exclusion chromatography with tandem multiangle laser light scattering and refractive index detection, was used to evaluate the kinetics of hydrolysis. The measurement of molecular weight provides a high degree of sensitivity in the case of small extents of reaction, thus allowing reliable measurements of the kinetics over short times. Pseudo-first-order rate constants for these polysaccharides were estimated using a simple model that accounts for the polydispersity of the starting sample. It was found that the relative order of backbone phosphodiester bond instability due to base hydrolysis was 19A > 10A > 19F > 6B > 17F, 20. Degradation of side-chain phosphodiester bonds was not observed, although the high degree of sensitivity in measurements is lost in this case, due to the low contribution of the side chains to the total polysaccharide molecular weight. In comparison with literature data on pneumococcal polysaccharide 6A, 19A was found to be the more labile, and hence appears to be the most labile pneumococcal polysaccharide studied to date. The rate of hydrolysis increased at higher pH and in the presence of divalent cation, but the extent was lower than expected based on similar data on RNA. Finally, the differences in the phosphodiester bond stabilities were analyzed by considering stereochemical factors in these polysaccharides. These results also provide a framework for evaluation of molecular integrity of phosphodiester-bond-containing polysaccharides in different solution conditions. Copyright 2004 Wiley Periodicals, Inc. Biopolymers, 2004
base hydrolysis, phosphodiester bond, pneumococcal polysaccharide
NCBI PubMed ID: 15307199Journal NLM ID: 0372525Publisher: Wiley Interscience
Correspondence: hari_pujar@merck.com
Institutions: Merck Research Laboratories, Merck & Co., West Point, PA 19486
- Article ID: 1013
Morona JK, Morona R, Paton JC "Characterization of the locus encoding the Streptococcus pneumoniae type 19F capsular polysaccharide biosynthetic pathway" -
Molecular Microbiology 23(4) (1997) 751-763
We have previously reported the nucleotide sequence of the first six genes of the Streptococcus pneumoniae type 19F capsular polysaccharide biosynthesis locus (cps19f). In this study we used plasmid insertion/rescue and inverse polymerase chain reaction (PCR) to clone an additional 10 kb downstream region containing the remainder of the cps19f locus, which was then subjected to sequence analysis. The cps19f locus is located in the S. pneumoniae chromosome between dexB and aliA, and consists of 15 open reading frames (ORFs), designated cps19fA to cps19fO, that appear to be arranged as a single transcriptional unit. Insertion-duplication mutants in seven out of the nine new ORFs have been constructed in a smooth type 19F strain, all of which resulted in a rough (nonencapsulated) phenotype, confirming that the operon is essential for capsule production. Comparison with sequence databases has allowed us to propose functions for 12 of the cps19f gene products, and a biosynthetic pathway for type 19F capsular polysaccharide. T7 expression studies confirmed that cps19fH, cps19fK, cps19fL, cps19fM and cps19fN directed the production of polypeptides of the expected size in Escherichia coli. The function of the cps19fK product was confirmed by its ability to complement a mutation in nfrC (rffE) in E. coli, as judged by restoration of sensitivity to bacteriophage N4. Interestingly, the last four genes of the locus (cps19fL-O) exhibit very strong homology (up to 70% amino acid identity) to a portion of the Shigella flexneri rfb gene cluster encoding biosynthesis of dTDP-rhamnose. When expressed in E. coli, cps19fL-O were capable of complementing a mutation deleting the respective Shigella flexneri homologues. Southern hybridization analysis indicated that cps19fA and cps19fB were the only cps genes found in all 16 S. pneumoniae serotypes/groups tested. The region from cps19fG to cps19fK was found only in members of serogroup 19, and, within this, cps19fl was unique to type 19F.
biosynthetic, capsular, characterization, polysaccharide, Streptococcus, Streptococcus pneumoniae, locus, capsular polysaccharide, type, pathway
NCBI PubMed ID: 9157246Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: patonj@wch.sa.gov.au
Institutions: Molecular Microbiology Unit, Woman's and Children's Hospital, North Adelaide, South Australia 5006, Australia, Departament of Microbiology ans Immunology, University of Adelaide, Adelaide, South Australia 5005, Australia.
- Article ID: 1307
Zamze S, Martinez-Pomares L, Jones H, Taylor PR, Stillion RJ, Gordon S, Wong SY "Recognition of bacterial capsular polysaccharides and lipopolysaccharides by the macrophage mannose receptor" -
Journal of Biological Chemistry 277(44) (2002) 41613-41623
The in vitro binding of the macrophage mannose receptor to a range of different bacterial polysaccharides was investigated. The receptor was shown to bind to purified capsular polysaccharides from Streptococcus pneumoniae and to the lipopolysaccharides, but not capsular polysaccharides, from Klebsiella pneumoniae. Binding was Ca(2+)- dependent and inhibitable with d-mannose. A fusion protein of the mannose receptor containing carbohydrate recognition domains 4-7 and a full-length soluble form of the mannose receptor containing all domains external to the transmembrane region both displayed very similar binding specificities toward bacterial polysaccharides, suggesting that domains 4-7 are sufficient for recognition of these structures. Surprisingly, no direct correlation could be made between polysaccharide structure and binding to the mannose receptor, suggesting that polysaccharide conformation may play an important role in recognition. The full-length soluble form of the mannose receptor was able to bind simultaneously both polysaccharide via the carbohydrate recognition domains and sulfated oligosaccharide via the cysteine-rich domain. The possible involvement of the mannose receptor, either cell surface or soluble, in the innate and adaptive immune responses to bacterial polysaccharides is discussed
lipopolysaccharides, structure, Streptococcus pneumoniae, capsular polysaccharides, recognition, Klebsiella pneumoniae, Mannose, macrophage, receptor
NCBI PubMed ID: 12196537Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: susanne.zamze@jenner.ac.uk
Institutions: Edward Jenner Institute for Vaccine Research, Compton, Berkshire RG20 7NN, United Kingdom and the Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom
- Article ID: 1347
Abeygunawardana C, Williams TC, Sumner JS, Hennessey JP "Development and validation of an NMR-based identity assay for bacterial polysaccharides" -
Analytical Biochemistry 279(2) (2000) 226-240
A method utilizing NMR spectroscopy has been developed to confirm the identity of bacterial polysaccharides used to formulate a polyvalent pneumococcal polysaccharide vaccine. The method is based on 600 MHz proton NMR spectra of individual serotype-specific polysaccharides. A portion of the anomeric region of each spectrum (5.89 to 4.64 ppm) is compared to spectra generated for designated reference samples for each polysaccharide of interest. The selected region offers a spectral window that is unique to a given polysaccharide and is sensitive to any structural alteration of the repeating units. The similarity of any two spectral profiles is evaluated using a correlation coefficient (rho), where rho >/= 0.95 between a sample and reference profile indicates a positive identification of the sample polysaccharide. This method has been shown to be extremely selective in its ability to discriminate between serotype-specific polysaccharides, some of which differ by no more than a single glycosidic linkage. Furthermore, the method is rapid and does not require extensive sample manipulations or pretreatments. The method was validated as a qualitative identity assay and will be incorporated into routine quality control testing of polysaccharide powders to be used in preparation of the polyvalent pneumococcal vaccine PNEUMOVAX 23. The specificity and reproducibility of the NMR-based identity assay is superior to the currently used colorimetric assays and can be readily adapted for use with other bacterial polysaccharide preparations as well.
NMR, Bacterial, polysaccharide, Bacterial polysaccharide, polysaccharides, bacterial polysaccharides, assay, development, identity assay, method development, validation
NCBI PubMed ID: 10706792Publication DOI: 10.1006/abio.1999.447Journal NLM ID: 0370535Publisher: Academic Press
Correspondence: abey@merck.com
Institutions: Bioprocess and Bioanalytical Research, Merck Research Laboratories, West Point, Pensylvania, USA
Methods: NMR
- Article ID: 1520
Jones C "NMR assays for carbohydrate-based vaccines" -
Journal of Pharmaceutical and Biomedical Analysis 38(5) (2005) 840-850
Antibodies against the cell surface carbohydrates of many microbial pathogens protect against infection. This was initially exploited by the development of purified polysaccharide vaccines, but glycoconjugate vaccines, in which the cell surface carbohydrate of a microbial pathogen is covalently attached to an appropriate carrier protein, are proving the most effective means to generate this protective immunity. Carbohydrate-based vaccines against Haemophilus influenzae Type b, Neisseria meningitidis, Streptococcus pneumoniae and Salmonella enterica serotype Typhi (S. Typhi) are already licensed, and many similar products are in various stages of development. For many of these vaccines, biological assays are not available or are inappropriate and NMR spectroscopy is proving a valuable tool for the characterisation and quality control of existing and novel products. This review highlights some of the areas in which NMR spectroscopy is currently used, and where further developments may be expected.
capsular polysaccharide, O-acetylation, pneumonia, glycoconjugate, meningitis, carbohydrate-based vaccines, identity, typhoid
NCBI PubMed ID: 16087046Publication DOI: 10.1016/j.jpba.2005.01.044Journal NLM ID: 8309336Publisher: London: Elsevier
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR
- Article ID: 1556
Pujar NS, Huang NF, Daniels CL, Dieter L, Gayton MG, Lee AL "Erratum: Base hydrolysis of phosphodiester bonds in pneumococcal polysaccharides" -
Biopolymers 77(6) (2005) 378-379
No abstract
polysaccharide, Streptococcus, Research, hydrolysis, polysaccharides, phosphodiester, pneumococcal, PDF, P, pneumococcal polysaccharides
NCBI PubMed ID: 15761954Journal NLM ID: 0372525Publisher: Wiley Interscience
Institutions: WP17-301, P. O. Box 4, Merck Research Laboratories, Merck & Co., West Point, PA 19486
- Article ID: 1919
Jennings HJ, Rosell KG, Carlo DJ "Structural determination of the capsular polysaccharide of Streptococcus pneumoniae type-19 (19F)" -
Canadian Journal of Chemistry 58(11) (1980) 1069-1074
The structure of the Pneumococcus type-19 (19F) capsular polysaccharide has been reinvestigated using nmr spectroscopy. It is composed of residues of 2-acetamido-2-deoxy-D-mannose, D-glucose, L-rhamnose, and phosphate in the molar ratio of 1:1:1:1. The polysaccharide is linear and is composed of the above components in a repeating unit of the following structure:→ 4)-β-D-ManNAcp-(1 → 4)-α-D-Glcp-(1→ 2)-α-L-Rhap-(1-PO4−)→.
Publication DOI: 10.1139/v80-167Journal NLM ID: 0372705Publisher: National Research Council of Canada Canada
Institutions: Division of Biological Sciiences, National Resreach Council of Canada, Ottawa, Ont., Canada KiA OR6
Methods: 13C NMR, 1H NMR, methylation, GLC-MS, gel filtration, partial acid hydrolysis, sugar analysis, GLC, optical rotation measurement
- Article ID: 1981
Ohno N, Yadomae T, Miyazaki T "The structure of the type-specific polysaccharide of Pneumococcus type XIX" -
Carbohydrate Research 80(2) (1980) 297-304
The structure of the capsular polysaccharide of Type XIX Streptococcus pneumoniae (S-XIX) has been elucidated by 1H- and 13C-n.m.r. spectroscopy. Mild hydrolysis of S-XIX with acid yielded a major oligosaccharide, the repeating unit of S-XIX, which was shown to be O-2-acetamido-2-deoxy-β-d-mannopyranosyl-(1→4)-O-α-d-glucopyranosyl-(1→2)-l-rhamnose 4′′-phosphate. Phosphoric acid forms a diester linkage in the S-XIX molecule, which explains the instability of S-XIX towards acid or alkali. The phosphodiester linkages in S-XIX join HO-1 of α-l-rhamnose and HO-4 of the 2-acetamido-2-deoxy-d-mannopyranosyl residue in the next repeating-unit. Treatment of S-XIX with alkali or alkaline-NaBH4 produced the repeating units in a lower yield. The proposed structure of S-XIX is [formula: see text].
Publication DOI: 10.1016/S0008-6215(00)84868-5Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Department of Microbial Chemistry, Tokyo College of Pharmacy, 1432-1 Horinouchi, Hachioji, Tokyo 192-03 Japan
Methods: 13C NMR, 1H NMR, gel filtration, mild acid hydrolysis, alkaline degradation, ion-exchange chromatography
- Article ID: 2251
Bednar B, Hennessey JP (JR) "Molecular size analysis of capsular polysaccharide preparations from Streptococcus pneumoniae" -
Carbohydrate Research 243 (1993) 115-130
Purified capsular polysaccharide preparations from Streptococcus pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were analyzed by high performance size exclusion chromatography (HPSEC) with multi-angle laser light scattering (MALLS), specific viscosity (SV), and refractive index (RI) detection to determine the molecular size and molar mass of each of the pneumococcal (Pn) polysaccharides. The Mw's of the polysaccharides ranged from a low of 606 kg/mol for Pn4 to a high of 1145 kg/mol for Pn9V, and the z-average radii of gyration ranged from 59 nm for Pn14 to 72 nm for Pn18C. Estimations of molar mass of the highly anionic polysaccharides (all but Pn14) by the universal calibration approach were unsuccessful, resulting in a 27-53% overestimate of the Mw's though application of Mark-Houwink-Sakurada coefficients calculated from the HPSEC-MALLS/SV/RI data resulted in estimates of Mw that were in agreement with the MALLS estimates for all but the Pn4 preparation. These results emphasize the need for direct measurement of both molecular size and intrinsic viscosity distributions for definitive characterization of the molar mass, hydrodynamic volume, rigidity, and drainage of complex biological polymers such as the pneumococcal polysaccharides.
NCBI PubMed ID: 8324758Publication DOI: 10.1016/0008-6215(93)84085-kJournal NLM ID: 0043535Publisher: Elsevier
Institutions: Department of Biological Chemistry, Merck Research Laboratories, West Point, Pennsylvania
- Article ID: 2515
Arndt B, Porro M "Strategies for type-specific glycoconjugate vaccines of Streptococcus pneumoniae" -
Advances in Experimental Medicine and Biology 303 (1991) 129-148
Journal NLM ID: 0121103Publisher: Kluwer Academic/Plenum Publishers
- Article ID: 2629
Lee CJ, Fraser BA "The structures of the cross-reactive types 19 (19F) and 57 (19A) pneumococcal capsular polysaccharides" -
Journal of Biological Chemistry 255 (1980) 6847-6853
Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
- Article ID: 3306
Guidolin A, Morona JK, Morona R, Hansman D, Paton JC "Nucleotide sequence analysis of genes essential for capsular polysaccharide biosynthesis in Streptococcus pneumoniae type 19F" -
Infection and Immunity 62(12) (1994) 5384-5396
Previous studies have shown that the capsular polysaccharide synthesis (cps) locus of the type 19F Streptococcus pneumoniae strain SSZ was closely linked to a copy of the insertion sequence IS1202 (J.K. Morona, A. Guidolin, R. Morona, D. Hansman, and J.C. Paton, J. Bacteriol. 176:4437-4443, 1994). In the present study, we used plasmid insertion and rescue and inverse PCR to clone 6,322 bp of flanking DNA upstream of IS1202. Sequence analysis indicated that this region contains six complete open reading frames (ORFs) and one partial ORF that are arranged as a single transcriptional unit. Chromosomal disruption of any of these ORFs in a smooth-type 19F strain leads to a rough (unencapsulated) phenotype, indicating that this operon is essential for capsule production. The ORFs have therefore been designated cps19fA to cps19fG, where cps19fA is the first gene of the type 19F cps locus. Furthermore, many of the gene products from this incomplete operon exhibit strong similarities to proteins known to be involved in the production of capsular polysaccharide, exopolysaccharide, teichoic acid, enterobacterial common antigen, and lipopolysaccharide from numerous other bacterial species. This has allowed us to propose functions for many of the type 19F cps gene products. Southern hybridization studies reveal that cps19fA and cps19fB are conserved among all 12 pneumococcal serotypes tested, whereas genes downstream of cps19fB are conserved among some, but not all, of the serotypes tested.
Lipopolysaccharide, biosynthesis, synthesis, antigen, common, gene, Bacterial, PCR, phenotype, Sequence Analysis, DNA, strain, capsular, polysaccharide, serotype, Streptococcus, Streptococcus pneumoniae, analysis, linked, locus, acid, conserved, capsular polysaccharide, type, protein, enterobacterial common antigen, exopolysaccharide, common antigen, enterobacterial, region, insertion, Open Reading Frames, production, lead, plasmid, rough, sequence, capsule, function, capsular polysaccharide biosynthesis, Serotypes, clone, teichoic acid, CPS, similarity, pneumococcal, proteins, operon, species, PDF, nucleotides, polysaccharide biosynthesis, sugar nucleotide, D
NCBI PubMed ID: 7960118Journal NLM ID: 0246127Publisher: American Society for Microbiology
Institutions: Department of Microbiology, Women's and Children's Hospital, North Adelaide, Australia.
Methods: genetic methods
- Article ID: 3365
Nikolaev AV, Botvinko IV, Ross AJ "Natural phosphoglycans containing glycosyl phosphate units: structural diversity and chemical synthesis" -
Carbohydrate Research 342(3-4) (2007) 297-344
An anomeric phosphodiester linkage formed by a glycosyl phosphate unit and a hydroxyl group of another monosaccharide is found in many glycopolymers of the outer membrane in bacteria (e.g., capsular polysaccharides and lipopolysaccharides), yeasts and protozoa. The polymers (phosphoglycans) composed of glycosyl phosphate (or oligoglycosyl phosphate) repeating units could be chemically classified as poly(glycosyl phosphates). Their importance as immunologically active components of the cell wall and/or capsule of numerous microorganisms upholds the need to develop routes for the chemical preparation of these biopolymers. In this paper, we (1) present a review of the primary structures (known to date) of natural phosphoglycans from various sources, which contain glycosyl phosphate units, and (2) discuss different approaches and recent achievements in the synthesis of glycosyl phosphosaccharides and poly(glycosyl phosphates).
synthesis, structure, polysaccharides, Phosphoglycans, Anomeric phosphodiesters
NCBI PubMed ID: 17092493Publication DOI: 10.1016/j.carres.2006.10.006Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: a.v.nikolaev@dundee.ac.uk
Institutions: College of Life Sciences, Division of Biological Chemistry and Molecular Microbiology, University of Dundee, Dundee DD1 5EH, UK.
- Article ID: 3666
Bonaccorsi F, Catelani G, Oscarson S "A new route for the synthesis of Streptococcus pneumoniae 19F and 19A capsular polysaccharide fragments avoiding the b-mannosamine glycosylation step" -
Carbohydrate Research 344(12) (2009) 1442-1448
The recently described [Attolino, E.; Bonaccorsi, F.; Catelani, G.; D'Andrea, F. Carbohydr. Res. 2008, 343, 2545-2556.] β-D-MaNAcp-(1→4)-β-D-Glcp thiophenyl glycosyl donor 3 was used in alpha-glycosylation reactions of OH-2 and OH-3 of the suitably protected p-MeO-benzyl α-L-rhamnopyranoside acceptors 7 and 8. Glycosylation of the axial OH-2 of 7 took place in high yield (76%) and with acceptable stereoselectivity (alpha/beta=3.4) leading to the protected trisaccharide alpha-11, corresponding to the repeating unit of Streptococcus pneumoniae 19F. The same reaction on equatorial OH-3 of acceptor 8 gave the trisaccharide alpha-15, a constituent of the repeating unit of S. pneumoniae 19A, but in lower yield (41%) and without stereoselection (alpha/beta=1:1.3). Utilizing the introduced orthogonal protection of OH-1 and OH-4'', the trisaccharide alpha-11 was transformed into a trisaccharide building block suitable for the synthesis of its phosphorylated oligomers.
Streptococcus pneumoniae, glycoconjugate vaccines, oligosaccharide synthesis, glycosylation, Thioglycosides
NCBI PubMed ID: 19467536Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: G. Catelani
; S.Oscarson
Institutions: Dipartimento di Chimica Bioorganica e Biofarmacia, Universita di Pisa, Via Bonanno 33, I-56126 Pisa, Italy, Centre for Synthesis and Chemical Biology, UCD School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland
Methods: 13C NMR, 1H NMR, chemical synthesis
- Article ID: 3753
Park S, Parameswar AR, Demchenko AV, Nahm MH "Identification of a simple chemical structure associated with protective human antibodies against multiple pneumococcal serogroups" -
Infection and Immunity 77(8) (2009) 3374-3379
Streptococcus pneumoniae, a major human pathogen, expresses at least 91 serologically distinct carbohydrate capsules. Since pneumococcal vaccines are designed to elicit antibodies against many different capsular polysaccharides (PSs), it is important to identify the epitopes involved in eliciting anti-capsular PS antibodies. We investigated the epitopes recognized by Dob1, which is a hybridoma-secreting human immunoglobulin G2 antibody to the PS of serotype 6B (Y. Sun et al., Infect. Immun. 67:1172-1179, 1999). We found that Dob1 bound synthetic capsular carbohydrates Gal(1→3)α-D-Glcp(1→3)α-L-Rhap(1→3)Rib-ol and α-D-Glcp(1→3)α-L-Rhap(1→3)Rib-ol but did not bind α-L-Rhap(1→3)Rib-ol. The critical epitope α-D-Glcp(1→3)α-L-Rhap is found in the capsular PSs of serotypes 6A, 6B, 6C, and 19A but not in the 19F PS. Consistent with this observation, Dob1 bound to the PSs of serotypes 6A, 6B, 6C, and 19A but did not bind the 19F PS and 23 additional unrelated pneumococcal capsular PSs. Also, Dob1 could opsonize pneumococci expressing serotypes 6A, 6B, 6C, and 19A but did not opsonize 19F pneumococci. In addition, ca. 7% of immune sera (12 of 175 sera) had significant amounts of Dob1-like antibodies, i.e., reacted with 6B and 19A PSs, but not with 19F PS. Humans can produce antibodies to the Dob1 epitope and the antibodies to that epitope cross-react with the four serotypes 6A, 6B, 6C, and 19A that belong to different serogroups. This epitope may be useful for producing a totally synthetic, simple chemical structure that is capable of generating protective antibodies to multiple pneumococcal serogroups.
Bacterial, Streptococcus pneumoniae, antibodies, Bacterial Capsule, sepitopes, Pneumococcal Infections
NCBI PubMed ID: 19451241Publication DOI: 10.1128/IAI.00319-09Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: nahm@uab.edu
Institutions: University of Alabama at Birmingham, 35294-2170, USA
Methods: ELISA, serological methods, IEF
- Article ID: 4431
Ovodov YS "Bacterial capsular antigens. Structural patterns of capsular antigens" -
Biochemistry (Moscow) 71(9) (2006) 937-954
Structural patterns of bacterial capsular antigens including capsular polysaccharides and exoglycans are given in this review. In addition, the immunological activity of capsular antigens and their role in type specificity of bacteria are discussed.
structure, capsular polysaccharides, bacterial capsular antigens, bacterial exoglycans, immunological activity, type specificity
NCBI PubMed ID: 17009947Publication DOI: 10.1134/S000629790609001XJournal NLM ID: 0376536Publisher: Nauka/Interperiodica
Correspondence: ovoys@physiol.komisc.ru
Institutions: Institute of Physiology, Komi Science Center, Urals Branch of the Russian Academy of Sciences, Syktyvkar 167982, Russia
- Article ID: 4695
Kuttel M, Gordon M, Ravenscroft N "Comparative simulation of pneumococcal serogroup 19 polysaccharide repeating units with two carbohydrate force fields" -
Carbohydrate Research 390 (2014) 20-27
Streptococcus pneumoniae causes meningitis, pneumonia and severe invasive disease (IPD) in young children. Although widespread infant immunisation with the PCV7 seven-valent pneumococcal conjugate vaccine has led to a dramatic decrease in IPD, infections due to non-vaccine serotypes, particularly serotype 19A, have increased. As the 19F polysaccharide differs from 19A at a single linkage position, it was assumed that PCV7 (containing 19F) would cross-protect against 19A disease. However, vaccination with PCV7 results in only 26% effectiveness against IPD caused by 19A. We explored the conformations and dynamics of the polysaccharide repeating units from serotypes 19F and 19A, comparing free energy surfaces for glycosidic linkages with 100ns aqueous molecular dynamics simulations of the di- and trisaccharide components. All calculations were performed with both the CHARMM and the GLYCAM carbohydrate force fields to establish whether the choice of model affects the predicted molecular behaviour. Although we identified key differences between the force fields, overall they were in agreement in predicting a 19F repeating unit with a wider range of conformation families than the more restricted 19A trisaccharide. This suggests a probable conformational difference between the 19F and 19A polysaccharides, which may explain the low cross-protection of 19F vaccines against 19A disease.
molecular modelling, conformation, chemistry, disease, invasive, polysaccharide, serotype, Streptococcus, Streptococcus pneumoniae, repeating unit, trisaccharide, carbohydrate, molecular, Research, glycosidic linkage, infection, conformational, dynamics, molecular dynamics, serogroup, linkage, polysaccharides, families, position, surface, vaccines, calculation, computer, difference, Serotypes, component, aqueous, vaccine, pneumococcal, simulation, conjugate, children, pneumonia, conjugate vaccine, model, decrease, free, force field, meningitis, energy, vaccination, pneumococcal conjugate vaccine, Science, cross-protection, effectiveness, Infant
Publication DOI: 10.1016/j.carres.2014.02.026Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mkuttel@cs.uct.ac.za (M.Kuttel)
Institutions: Department of Computer Science, University of Cape Town, Cape Town, South Africa
Methods: conformation analysis, MD simulations, molecular modeling, MM3 force field
- Article ID: 4768
Kuttel MM, Jackson GE, Mafata M, Ravenscroft N "Capsular polysaccharide conformations in pneumococcal serotypes 19F and 19A" -
Carbohydrate Research 406 (2015) 27-33
Streptococcus pneumoniae is a significant pathogen in children. Although the PCV7 pneumococcal conjugate vaccine has reduced pneumococcal disease, non-vaccine serotype 19A infection has increased, despite expectations of cross-protection from vaccine serotype 19F. Serotype 19A is included in the new PCV13 vaccine, but not in PCV10. In the solution simulations of 19F and 19A oligosaccharide chains reported here, both polysaccharides form unstructured random coils, with inflexible repeat units linked by mobile phosphodiester linkages. However, there are clear conformational differences. In the 19F repeat unit, the rhamnose residue is nearly orthogonal to the other residues, whereas 19A has residues in similar orientations. This finding is corroborated by key inter-residue distances calculated from NMR NOESY experiments. Further, 19F is predominantly in extended conformations, whereas 19A exhibits a high prevalence of tight hairpin bends. These conformational differences may account for a lack of antibody cross-protection between serotypes 19F and 19A.
molecular modelling, Streptococcus pneumoniae, vaccine, pneumococcal, cross protection
NCBI PubMed ID: 25658063Publication DOI: 10.1016/j.carres.2014.12.013Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mkuttel@cs.uct.ac.za
Institutions: Department of Computer Science, University of Cape Town, Cape Town, South Africa
Methods: 13C NMR, 1H NMR, NMR-2D, conformation analysis, molecular modeling, MD simulation, polymer hydrodynamics
- Article ID: 4828
Geno KA, Gilbert GL, Song JY, Skovsted IC, Klugman KP, Jones C, Konradsen HB, Nahm MH "Pneumococcal Capsules and Their Types: Past, Present, and Future" -
Clinical Microbiology Reviews 28(3) (2015) 871-899
Streptococcus pneumoniae (the pneumococcus) is an important human pathogen. Its virulence is largely due to its polysaccharide capsule, which shields it from the host immune system, and because of this, the capsule has been extensively studied. Studies of the capsule led to the identification of DNA as the genetic material, identification of many different capsular serotypes, and identification of the serotype-specific nature of protection by adaptive immunity. Recent studies have led to the determination of capsular polysaccharide structures for many serotypes using advanced analytical technologies, complete elucidation of genetic basis for the capsular types, and the development of highly effective pneumococcal conjugate vaccines. Conjugate vaccine use has altered the serotype distribution by either serotype replacement or switching, and this has increased the need to serotype pneumococci. Due to great advances in molecular technologies and our understanding of the pneumococcal genome, molecular approaches have become powerful tools to predict pneumococcal serotypes. In addition, more-precise and -efficient serotyping methods that directly detect polysaccharide structures are emerging. These improvements in our capabilities will greatly enhance future investigations of pneumococcal epidemiology and diseases and the biology of colonization and innate immunity to pneumococcal capsules.
serotype, Streptococcus pneumoniae, vaccines, Pneumococcal Capsules
NCBI PubMed ID: 26085553Publication DOI: 10.1128/CMR.00024-15Journal NLM ID: 8807282Publisher: Washington, DC: American Society for Microbiology
Correspondence: Moon H. Nahm
Institutions: Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama, USA, Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology & Medical Research, Westmead Hospital, Wentworthville, New South Wales, Australia, Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Sydney, New South Wales, Australia, Division of Infectious Disease, Department of Internal Medicine, Korea University Guro Hospital, Seoul, South Korea, SSI Diagnostica, Division of Microbiology and Diagnostics, Statens Serum Institut, Copenhagen, Denmark, Pneumonia Program Strategy Team, Bill & Melinda Gates Foundation, Seattle, Washington, USA, Laboratory for Molecular Structure, NIBSC, South Mimms, Herts, United Kingdom, Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, USA
- Article ID: 4836
Berti F, Ravenscroft N "Characterization of Carbohydrate Vaccines by NMR Spectroscopy" -
Methods in Molecular Biology 1331 (2015) 189-209
Physicochemical techniques are a powerful tool for the structural characterization of carbohydrate-based vaccines. High-field Nuclear Magnetic Resonance (NMR) spectroscopy has been established as an extremely useful and robust method for tracking the industrial manufacturing process of these vaccines from polysaccharide bulk antigen through to the final formulation. Here, we describe the use of proton NMR for structural identity and conformity testing of carbohydrate-based vaccines.
carbohydrates, capsular polysaccharide, antigens, nuclear magnetic resonance spectroscopy, vaccines
NCBI PubMed ID: 26169742Publication DOI: 10.1007/978-1-4939-2874-3_12Journal NLM ID: 9214969Publisher: Springer
Correspondence: francesco.x.berti@gsk.com
Institutions: Research, GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy
- Article ID: 5440
Goyette-Desjardins G, Vinogradov E, Okura M, Takamatsu D, Gottschalk M, Segura M "Structure determination of Streptococcus suis serotypes 7 and 8 capsular polysaccharides and assignment of functions of the cps locus genes involved in their biosynthesis" -
Carbohydrate Research 473 (2019) 36-45
Streptococcus suis serotypes 7 and 8 are counted among the top six S. suis serotypes causing clinical disease in pigs. Yet, limited information is available on these serotypes. Since S. suis serotyping system is based upon capsular polysaccharide (CPS) antigenicity and the CPS is considered a major virulence factor for encapsulated pathogens, here we determined for the first time the chemical compositions and structures of serotypes 7 and 8 CPSs. Chemical and spectroscopic data gave the following repeating unit sequences: [3)L-Rha(α1-P-2)D-Gal(α1-4)D-GlcA(β1-3)D-FucNAc4N(α1-]n for serotype 7 and [2)L-Rha(α1-P-4)D-ManNAc(β1-4)D-Glc(α1-]n for serotype 8. As serotype 8 CPS is identical to Streptococcus pneumoniae type 19F CPS, dot-blot analyses showed a strong reaction of the 19F polysaccharide with reference anti-S. suis serotype 8 rabbit serum. A correlation between S. suis serotypes 7 and 8 sequences and genes of those serotypes' loci encoding putative glycosyltransferases and polymerases responsible for the biosynthesis of the repeating units was tentatively established. Knowledge of CPS structure and composition will contribute to better dissect the role of this bacterial component in the pathogenesis of the disease caused by S. suis serotypes 7 and 8.
capsular polysaccharide, carbohydrate structure, Serotype 8, Streptococcus suis, Serotype 7
NCBI PubMed ID: 30605786Publication DOI: 10.1016/j.carres.2018.12.009Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: M. Segura
Institutions: Swine and Poultry Infectious Diseases Research Center, Faculty of Veterinary Medicine, University of Montreal, 3200 Sicotte St., St-Hyacinthe, Quebec, J2S 2M2, Canada, Canadian Glycomics Network (GlycoNet), University of Alberta, 11227 Saskatchewan Dr., Edmonton, Alberta, T6G 2G2, Canada, National Research Council, 100 Sussex Dr., Ottawa, Ontario, K1A 0R6, Canada, Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan, The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu, Gifu, 501-1193, Japan
Methods: 13C NMR, 1H NMR, GLC-MS, gel filtration, NMR-2D, sugar analysis, 31P NMR, acid hydrolysis, Western blotting, bioinformatic analysis
- Article ID: 5473
Zou W, Li J, Vinogradov E, Cox A "Removal of cell wall polysaccharide in pneumococcal capsular polysaccharides by selective degradation via deamination" -
Carbohydrate Polymers 218 (2019) 199-207
Pneumococcal cell wall polysaccharide (C-PS), a contaminant in pneumococcal capsular polysaccharide (Pn-PS) vaccines is degraded by mild deamination of the 4-amino-2-acetamido-2,4,6-tri-deoxy-galactose (AAT) in C-PS, which was carried out by addition of 5% aqueous sodium nitrite to a solution of polysaccharide in 5% aqueous acetic acid. Glycosidic linkage and functional groups such as O-acetates, phosphodiesters, and pyruvates were preserved under the conditions. The small fragments from degraded C-PS were removed by ultrafiltration or dialysis to provide essentially C-PS free Pn-PS. Because of the presence of AAT in its structure the deamination is not suitable for the purification of type 1 Pn-PS. Meanwhile, the mass and NMR spectroscopic analysis on the deamination products suggests that both type 1 Pn-PS and C-PS degraded following a major pathway of 5,4-hydride shift, cleavage of AAT O5-C1 bond, C1 hemiacetal formation, and its hydrolysis to release neighboring GalA- in type 1 Pn-PS and GalNAc(6-O-PCho)- in C-PS
mechanism, degradation, deamination, cell wall polysaccharide, pneumococcal capsular polysaccharide
NCBI PubMed ID: 31221321Publication DOI: 10.1016/j.carbpol.2019.03.070Journal NLM ID: 8307156Publisher: Elsevier
Correspondence: W. Zou
Institutions: Human Health Therapeutic Research Center, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, K1A 0R6, Canada
Methods: 13C NMR, 1H NMR, gel filtration, sugar analysis, MS/MS, MS, dialysis, SEC-HPLC, ultrafiltration, mild deamination
- Article ID: 5776
Goyette-Desjardins G, Auger JP, Dolbec D, Vinogradov E, Okura M, Takamatsu D, Van Calsteren M, Gottschalk M, Segura M "Comparative Study of Immunogenic Properties of Purified Capsular Polysaccharides from Streptococcus suis Serotypes 3, 7, 8, and 9: the Serotype 3 Polysaccharide Induces an Opsonizing IgG Response" -
Infection and Immunity 88(10) (2020) e00377
Streptococcus suis is an encapsulated bacterium and one of the most important swine pathogens and a zoonotic agent for which no effective vaccine exists. Bacterial capsular polysaccharides (CPSs) are poorly immunogenic, but anti-CPS antibodies are essential to the host defense against encapsulated bacteria. In addition to the previously known serotypes 2 and 14, which are nonimmunogenic, we have recently purified and described the CPS structures for serotypes 1, 1/2, 3, 7, 8, and 9. Here, we aimed to elucidate how these new structurally diverse CPSs interact with the immune system to generate anti-CPS antibody responses. CPS-stimulated dendritic cells produced significant levels of C-C motif chemokine ligand 3 (CCL3), partially via Toll-like receptor 2 (TLR2)- and myeloid differentiation factor 88-dependent pathways, and CCL2, via TLR-independent mechanisms. Mice immunized with purified serotype 3 CPS adjuvanted with TiterMax Gold produced an opsonizing IgG response, whereas other CPSs or adjuvants were negative. Mice hyperimmunized with heat-killed S. suis serotypes 3 and 9 both produced anti-CPS type 1 IgGs, whereas serotypes 7 and 8 remained negative. Also, mice infected with sublethal doses of S. suis serotype 3 produced primary anti-CPS IgM and IgG responses, of which only IgM were boosted after a secondary infection. In contrast, mice sublethally infected with S. suis serotype 9 produced weak anti-CPS IgM and IgG responses following a secondary infection. This study provides important information on the divergent evolution of CPS serotypes with highly different structural and/or biochemical properties within S. suis and their interaction with the immune system.
capsular polysaccharide, immunogenicity, Streptococcus suis, Serotype 3
NCBI PubMed ID: 32747605Publication DOI: 10.1128/IAI.00377-20Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: Mariela Segura
Institutions: National Research Council, Ottawa, Ontario, Canada, The United Graduate School of Veterinary Sciences, Gifu University, Gifu, Gifu, Japan, Canadian Glycomics Network (GlycoNet), University of Alberta, Edmonton, AB, Canada, Swine and Poultry Infectious Diseases Research Centre, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, Quebec, Canada, Research Group on Infectious Diseases in Production Animals, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, Quebec, Canada, Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Japan
Methods: NanoOrange assay, chemokine production, immunogenicity studies
- Article ID: 5791
Knirel YA, Van Calsteren M "Bacterial exopolysaccharides" -
Book: Comprehensive Glycoscience: From Chemistry to Systems Biology. Reference Module in Chemistry, Molecular Sciences and Chemical Engineering (2021) 1-75
Bacterial extracellular polysaccharides are known as a cell-bound capsule, a sheath, or a slime, which is excreted into the environment. They play an important role in virulence of medical bacteria and plant-to-symbiont interaction and are used for serotyping of bacteria and production of vaccines. Some exopolysaccharides have commercial applications in industry, and claims of health benefits have been documented for an increasing number of them. Exopolysaccharides have diverse composition and structure, and some contain sugar and non-sugar components that are found in bacterial carbohydrates only. The present article provides an updated collection of the data on exopolysaccharides of various classes of gram-negative and gram-positive bacteria reported until the end of 2019. When known, biosynthesis pathways of exopolysaccharides are treated in a summary manner. References are made to structure and biosynthesis relatedness between exopolysaccharides of different bacterial taxa as well as between bacterial polysaccharides and mammalian glycosaminoglycans.
polysaccharide structure, Gram-negative bacteria, capsule, Biofilm, polysaccharide biosynthesis, gram-positive bacteria, Monosaccharide composition, Bacterial exopolysaccharide, non-sugar component
Publication DOI: 10.1016/B978-0-12-819475-1.00005-5Publisher: Elsevier
Correspondence: marie-rose.vancalsteren@canada.ca; yknirel@gmail.com
Editors: Barchi J, Kamerling H
Institutions: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Saint-Hyacinthe Research and Development Centre, Agriculture and Agri-Food Canada, Saint-Hyacinthe, QC, Canada
- Article ID: 6058
Genning M, Kurbatova EA, Nifantiev NE "Synthetic Analogs of Streptococcus pneumoniae Capsular Polysaccharides and Immunogenic Activities of Glycoconjugates" -
Russian Journal of Bioorganic Chemistry 47(1) (2021) 1-25
treptococcus pneumoniae is a Gram-positive bacterium (pneumococcus) that causes severe diseases in adults and children. It was established that some capsular polysaccharides of the clinically significant serotypes of S. pneumoniae in the composition of commercial pneumococcal polysaccharide or conjugate vaccines exhibit low immunogenicity. The review considers production methods and structural features of the synthetic oligosaccharides from the problematic pneumococcal serotypes that are characterized with low immunogenicity due to destruction or detrimental modification occurring in the process of their preparation and purification. Bacterial serotypes that cause severe pneumococcal diseases as well as serotypes not included in the composition of the pneumococcal conjugate vaccines are also discussed. It is demonstrated that the synthetic oligosaccharides corresponding to protective glycotopes of the capsular polysaccharides of various pneumococcal serotypes are capable of inducing formation of the protective opsonizing antibodies and immunological memory. Optimal constructs of oligosaccharides from the epidemiologically significant pneumococcal serotypes are presented that can be used for designing synthetic pneumococcal vaccines, as well as test systems for diagnosis of S. pneumoniae infections and monitoring of vaccination efficiency.
oligosaccharide, antibodies, ligand, vaccine, immunogen, Opsonophagocytosis, protective activity, pneumococci
NCBI PubMed ID: 33776393Publication DOI: 10.1134/S1068162021010076Journal NLM ID: 9420101Publisher: Springer Science and Business Media
Correspondence: nen@ioc.ac.ru
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Mechnikov Research Institute for Vaccines and Sera, 105064 Moscow, Russia
- Article ID: 6073
Javed J, Mandal PK "Bacterial surface capsular polysaccharides from Streptococcus pneumoniae: A systematic review on structures, syntheses, and glycoconjugate vaccines" -
Carbohydrate Research 502 (2021) 108277
The polysaccharide capsule of Streptococcus pneumoniae constitutes the outermost surface structure of the organism and plays a critical role in virulence. The capsule is the target of current pneumococcal vaccines and glycoconjugates and has important medical and industrial applications. Widespread use of these vaccines is driving changes in serotype prevalence in disease. A massive array of sugars and glycosidic linkages experienced with complete diversity of potential polysaccharide structures. However, it is impossible to collect a sufficient quantity of glycan antigens for the preparation of CPS-based glycoconjugate vaccines from natural sources with high purity and for thorough biological evaluation. So nowadays, the development of a chemical synthetic strategy and their conjugation with a carrier protein to form synthetic glycoconjugate vaccines has been used to gain access on a large scale. This review provides a comprehensive summary of structures, synthesis as well as recent development of synthetic glycoconjugate vaccines, which will support research and may benefit the glycochemical and medical sciences.
synthesis, Streptococcus pneumoniae, glycoconjugate vaccines, polysaccharides, Pneumococcal Infections
NCBI PubMed ID: 33743443Publication DOI: 10.1016/j.carres.2021.108277Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: P.K. Mandal
Institutions: Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, BS-10/1, Sector 10, Jankipuram Extension, Sitapur Road, P.O. Box 173, Lucknow, 226 031, India, Academy of Scientific and Innovative Research (AcSIR), New Delhi, India
- Article ID: 6114
Morelli L, Lay L, Santana-Mederos D, Valdes-Balbin Y, Verez Bencomo V, van Diepen A, Hokke CH, Chiodo F, Compostella F "Glycan Array Evaluation of Synthetic Epitopes between the Capsular Polysaccharides from Streptococcus pneumoniae 19F and 19A" -
ACS Chemical Biology 16(9) (2021) 1671-1679
Vaccination represents the most effective way to prevent invasive pneumococcal diseases. The glycoconjugate vaccines licensed so far are obtained from capsular polysaccharides (CPSs) of the most virulent serotypes. Protection is largely limited to the specific vaccine serotypes, and the continuous need for broader coverage to control the outbreak of emerging serotypes is pushing the development of new vaccine candidates. Indeed, the development of efficacious vaccine formulation is complicated by the high number of bacterial serotypes with different CPSs. In this context, to simplify vaccine composition, we propose the design of new saccharide fragments containing chemical structures shared by different serotypes as cross-reactive and potentially cross-protective common antigens. In particular, we focused on Streptococcus pneumoniae (Sp) 19A and 19F. The CPS repeating units of Sp 19F and 19A are very similar and share a common structure, the disaccharide ManNAc-β-(1→4)-Glc (A-B). Herein, we describe the synthesis of a small library of compounds containing different combinations of the common 19F/19A disaccharide. The six new compounds were tested with a glycan array to evaluate their recognition by antibodies in reference group 19 antisera and factor reference antisera (reacting against 19F or 19A). The disaccharide A-B, phosphorylated at the upstream end, emerged as a hit from the glycan array screening because it is strongly recognized by the group 19 antisera and by the 19F and 19A factor antisera, with similar intensity compared with the CPSs used as controls. Our data give a strong indication that the phosphorylated disaccharide A-B can be considered a common epitope among different Sp 19 serotypes.
Streptococcus pneumoniae, capsular polysaccharides, glycan, disaccharide, glycoconjugate vaccine, vaccination, glycan epitopes
NCBI PubMed ID: 34469105Publication DOI: 10.1021/acschembio.1c00347Journal NLM ID: 101282906Publisher: Washington, DC: American Chemical Society
Correspondence: Fabrizio Chiodo < f.chiodo@amsterdamumc.nl>; Federica Compostella
Institutions: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Saldini 50, 20133 Milano, Italy, Department of Chemistry, University of Milan, Via Golgi 19, 20133 Milano, Italy, Finlay Vaccine Institute, 200 and 21 Street, 11600 Havana, Cuba, Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands, Italian National Research Council (CNR), Institute of Biomolecular Chemistry (ICB), Via Campi Flegrei 34, 80078 Pozzuoli, Italy
Methods: 13C NMR, 1H NMR, NMR-2D, TLC, 31P NMR, ESI-MS, chemical synthesis, chemical methods, immunization, glycan array analysis, immobilization
- Article ID: 6118
Okura M, Auger JP, Shibahara T, Goyette-Desjardins G, Van Glsteren MR, Maruyama F, Kawai M, Osaki M, Segura M, Gottschalk M, Takamatsu D "Capsular polysaccharide switching in Streptococcus suis modulates host cell interactions and virulence" -
Scientific Reports 11(1) (2021) 6513
The capsular polysaccharide (CPS) of Streptococcus suis defines various serotypes based on its composition and structure. Though serotype switching has been suggested to occur between S. suis strains, its impact on pathogenicity and virulence remains unknown. Herein, we experimentally generated S. suis serotype-switched mutants from a serotype 2 strain that express the serotype 3, 4, 7, 8, 9, or 14 CPS. The effects of serotype switching were then investigated with regards to classical properties conferred by presence of the serotype 2 CPS, including adhesion to/invasion of epithelial cells, resistance to phagocytosis by macrophages, killing by whole blood, dendritic cell-derived pro-inflammatory mediator production and virulence using mouse and porcine infection models. Results demonstrated that these properties on host cell interactions were differentially modulated depending on the switched serotypes, although some different mutations other than loci of CPS-related genes were found in each the serotype-switched mutant. Among the serotype-switched mutants, the mutant expressing the serotype 8 CPS was hyper-virulent, whereas mutants expressing the serotype 3 or 4 CPSs had reduced virulence. By contrast, switching to serotype 7, 9, or 14 CPSs had little to no effect. These findings suggest that serotype switching can drastically alter S. suis virulence and host cell interactions.
disease, Bacterial, virulence, serotype, capsular polysaccharide, Streptococcus suis, animal health
NCBI PubMed ID: 33753801Publication DOI: 10.1038/s41598-021-85882-3Journal NLM ID: 101563288Publisher: London: Nature Publishing Group
Correspondence: mokura@affrc.go.jp; mariela.segura@umontreal.ca; marcelo.gottshcalk@umontreal.ca
Institutions: The United Graduate School of Veterinary Sciences, Gifu University, Gifu, Gifu, Japan, Division of Bacterial and Parasitic Diseases, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Japan, Saint-Hyacinthe Research and Development Centre, Agriculture and Agri-Food Canada, Saint-Hyacinthe, QC, Canada, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada, Division of Pathology and Pathophysiology, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Japan, Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan, Microbial Genomics and Ecology, Office of Industry-Academia-Government and Community Collaboration, Hiroshima University, Hiroshima, Japan, Scientific and Technological Bioresource Nucleus, Universidad de La Frontera, Temuco, Chile, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto, Japan
Methods: 13C NMR, 1H NMR, NMR-2D, PCR, ELISA, genetic methods, serotyping, bactericidal assays, TEM, phagocytosis assay, genome sequencing
- Article ID: 6178
Zhang S, Sella M, Sianturi J, Priegue P, Shen D, Seeberger PH "Discovery of Oligosaccharide Antigens for Semi-Synthetic Glycoconjugate Vaccine Leads against Streptococcus suis Serotypes 2, 3, 9 and 14" -
Angewandte Chemie, International Edition 60(26) (2021) 14679-14692
Streptococcus suis bacteria are one of the most serious health problems for pigs and an emerging zoonotic agent in humans working in the swine industry. S. suis bacteria express capsular polysaccharides (CPS) a major bacterial virulence factor that define the serotypes. Oligosaccharides resembling the CPS of S. suis serotypes 2, 3, 9, and 14 have been synthesized, glycans related to serotypes 2 and 9 were placed on glycan array surfaces to screen blood from infected pigs. Lead antigens for the development of semi-synthetic S. suis serotypes 2 and 9 glycoconjugate veterinary vaccines were identified in this way.
carbohydrates, Oligosaccharides, immunology, glycans, total synthesis, Streptococcus suis
NCBI PubMed ID: 33852172Publication DOI: 10.1002/anie.202103990Journal NLM ID: 0370543Publisher: Weinheim: Wiley-VCH
Correspondence: Peter.Seeberger@mpikg.mpg.de
Institutions: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195, Berlin, Germany, Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Muhlenberg 1, 14476, Potsdam, Germany, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA, 02138, USA
Methods: 13C NMR, 1H NMR, NMR-2D, TLC, ELISA, chemical synthesis, chemical methods, FTIR, HPLC, glycosylation, optical rotation measurement, glycan array analysis, HR-ESI-MS, flash chromatography
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10. Compound ID: 825
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_149136,IEDB_151531,IEDB_885813
The structure is contained in the following publication(s):
- Article ID: 226
Winn AM, Wilkinson SG "Structure of the O16 antigen of Stenotrophomonas maltophilia" -
Carbohydrate Research 330(2) (2001) 279-283
A polysaccharide containing D-ribose, N-acetyl-D-glucosamine, and N-acetyl-D-mannosamine was isolated from the phenol-soluble lipopolysaccharide extracted from defatted cell walls of the reference strain (560) for serogroup O16 of Stenotrophomonas maltophilia. The results of methylation analysis, chemical degradations, and NMR spectroscopy showed that the polysaccharide is based on a branched trisaccharide repeating-unit of the structure shown below. Although ribose was absent from about half of the units in the isolated polymer, the regularity and spacing of the ladder observed on SDS-PAGE of the parent lipopolysaccharide indicate that this was an artefact of the mild acid hydrolysis used to release the polymer. On the other hand, the effects of mild alkaline hydrolysis on the polymer indicated partial O-acetylation.
Lipopolysaccharide, Stenotrophomonas maltophilia, O-specific polymer
NCBI PubMed ID: 11217982Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: s.g.wilkinson@chem.hull.ac.uk
Institutions: Department of Chemistry, Faculty of Science and the Environment, University of Hull, Hull HU6 7RX, UK
Methods: methylation, NMR, chemical degradation
- Article ID: 4329
Knirel YA "Structure of O-antigens" -
Book: Bacterial lipopolysaccharides: Structure, chemical synthesis, biogenesis and interaction with host cells (2011) Chapter 3, 41-115
The lipopolysaccharide (LPS) is the major constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. Its lipid A moiety is embedded in the membrane and serves as an anchor for the rest of the LPS molecule. The outermost repetitive glycan region of the LPS is linked to the lipid A through a core oligosaccharide (OS), and is designated as the O-specific polysaccharide (O-polysaccharide, OPS) or O-antigen. The O-antigen is the most variable portion of the LPS and provides serological specificity, which is used for bacterial serotyping. The OPS also provides protection to the microorganisms from host defenses such as complement mediated killing and phagocytosis, and is involved in interactions of bacteria with plants and bacteriophages. Studies of the OPSs ranging from the elucidation of their chemical structures and conformations to their biological and physico-chemical properties help improving classification schemes of Gram-negative bacteria. Furthermore, these studies contributed to a better understanding of the mechanisms of pathogenesis of infectious diseases, as well as provided information to develop novel vaccines and diagnostic reagents.
Lipopolysaccharide, synthesis, lipopolysaccharides, structure, Bacterial, host, O-antigen, O antigen, cell, O antigens, O-antigens, chemical, interaction, cells, PDF, chemical synthesis, biogenesis
Publication DOI: 10.1007/978-3-7091-0733-1_3Publisher: Springer
Correspondence: knirel@ioc.ac.ru
Editors: Knirel YA, Valvano MA
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
Expand this compound
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11. Compound ID: 826
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, cell wall polysaccharide
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_885813
The structure is contained in the following publication(s):
- Article ID: 226
Winn AM, Wilkinson SG "Structure of the O16 antigen of Stenotrophomonas maltophilia" -
Carbohydrate Research 330(2) (2001) 279-283
A polysaccharide containing D-ribose, N-acetyl-D-glucosamine, and N-acetyl-D-mannosamine was isolated from the phenol-soluble lipopolysaccharide extracted from defatted cell walls of the reference strain (560) for serogroup O16 of Stenotrophomonas maltophilia. The results of methylation analysis, chemical degradations, and NMR spectroscopy showed that the polysaccharide is based on a branched trisaccharide repeating-unit of the structure shown below. Although ribose was absent from about half of the units in the isolated polymer, the regularity and spacing of the ladder observed on SDS-PAGE of the parent lipopolysaccharide indicate that this was an artefact of the mild acid hydrolysis used to release the polymer. On the other hand, the effects of mild alkaline hydrolysis on the polymer indicated partial O-acetylation.
Lipopolysaccharide, Stenotrophomonas maltophilia, O-specific polymer
NCBI PubMed ID: 11217982Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: s.g.wilkinson@chem.hull.ac.uk
Institutions: Department of Chemistry, Faculty of Science and the Environment, University of Hull, Hull HU6 7RX, UK
Methods: methylation, NMR, chemical degradation
- Article ID: 3112
Kojima N, Kaya S, Araki Y, Ito E "Pyruvic-acid-containing polysaccharide in the cell wall of Bacillus polymyxa AHU 1385" -
European Journal of Biochemistry 174 (1988) 255-260
Three acidic polymer fractions with molecular masses of about 16 kDa, 35 kDa and 70 kDa were isolated from lysozyme digests of N-acetylated cell walls of Bacillus polymyxa AHU 1385 by ion-exchange chromatography and gel chromatography. These fractions, containing mannosamine, glucosamine and pyruvic acid in a molar ratio of about 1:1:1 together with glycopeptide components, were characterized as polysaccharide-linked glycopeptides with one, two and more polysaccharide chains. On the other hand, treatment of the cell walls with glycine/HC1 buffer, pH 2.5, at 100 degrees C for 10 min followed by separation of water-soluble products on ion-exchange chromatography gave three polysaccharide fractions, PS-I-III, which contained different amounts of pyruvic acid (0,0.6 and 0.9 residue/mannosamine residue) along with equimolar amounts of mannosamine and glucosamine. Pyruvate-free polysaccharides similar to PS-I were also obtained from PS-II, PS-III and polysaccharide-linked glycopeptides by treatment with 10 mM HC1 at 100 degrees C for 1 h. Results of analyses of these polysaccharide preparations by 1H-NMR and 13C-NMR measurement and methylation, together with data from characterization of fragments obtained by hydrogen fluoride hydrolysis, lead to the most likely structure, →3)[4,6-O-(1-carboxyethylidene)]ManNAc(β1→4)GlcNac(β1→, for the acidic polysaccharide of this strain.
NCBI PubMed ID: 3383845Publication DOI: 10.1111/j.1432-1033.1988.tb14091.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Chemistry, Faculty of Science, Hokkaido University
Methods: 13C NMR, 1H NMR, GC-MS
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12. Compound ID: 833
| Cyclic
-3)-a-D-Fucp4NAc-(1-4)-b-D-ManpNAc-(1-4)-a-D-GlcpNAc-(1- |
Show graphically |
Structure type: cyclic polymer repeating unit
; n=4, 2430
Trivial name: Enterobacterial common antigen
Contained glycoepitopes: IEDB_137340,IEDB_141807,IEDB_151531,IEDB_885813
The structure is contained in the following publication(s):
- Article ID: 230
Erbel PJ, Barr K, Gao N, Gerwig GJ, Rick PD, Gardner KH "Identification and biosynthesis of cyclic enterobacterial common antigen in Escherichia coli" -
Journal of Bacteriology 185(6) (2003) 1995-2004
Phosphoglyceride-linked enterobacterial common antigen (ECA(PG)) is a cell surface glycolipid that is synthesized by all gram-negative enteric bacteria. The carbohydrate portion of ECA(PG) consists of linear heteropolysaccharide chains comprised of the trisaccharide repeat unit Fuc4NAc-ManNAcA-GlcNAc, where Fuc4NAc is 4-acetamido-4,6-dideoxy-D-galactose, ManNAcA is N-acetyl-D-mannosaminuronic acid, and GlcNAc is N-acetyl-D-glucosamine. The potential reducing terminal GlcNAc residue of each polysaccharide chain is linked via phosphodiester linkage to a phosphoglyceride aglycone. We demonstrate here the occurrence of a water-soluble cyclic form of enterobacterial common antigen, ECA(CYC), purified from Escherichia coli strains B and K-12 with solution nuclear magnetic resonance (NMR) spectroscopy, electrospray ionization mass spectrometry (ESI-MS), and additional biochemical methods. The ECA(CYC) molecules lacked an aglycone and contained four trisaccharide repeat units that were nonstoichiometrically substituted with up to four O-acetyl groups. ECA(CYC) was not detected in mutant strains that possessed null mutations in the wecA, wecF, and wecG genes of the wec gene cluster. These observations corroborate the structural data obtained by NMR and ESI-MS analyses and show for the first time that the trisaccharide repeat units of ECA(CYC) and ECA(PG) are assembled by a common biosynthetic pathway.
NMR, biosynthesis, antigen, common, chemistry, Bacterial, genetics, growth & development, metabolism, potential, strain, Non-U.S.Gov't, terminal, polysaccharide, trisaccharide, carbohydrate, cell, chain, linked, form, Escherichia, Escherichia coli, acid, antigens, immunology, enterobacterial common antigen, bacteria, electrospray, spectrometry, linkage, common antigen, cyclic, enterobacterial, identification, biochemistry, Magnetic Resonance Spectroscopy, nuclear, nuclear magnetic resonance, resonance, spectroscopy, surface, Gram-negative, mutation, purified, methods, glycolipid, reducing, heteropolysaccharide, linear, occurrence, phosphodiester, solution, Enterobacteriaceae, U.S.Gov't, electrophoresis, isolation & purification, Mass, Electrospray Ionization, P.H.S., Research Support, Basic Helix-Loop-Helix Transcription Factors, Trans-Activators, Transcription Factors
NCBI PubMed ID: 12618464Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: Kevin.Gardner@UTSouthwestern.edu
Institutions: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9038, USA, Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9038, Department of Microbiology and Immunology, F. Edward Herbert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, Maryland 208142, Department of Bio-Organic Chemistry, Bijvoet Center, Utrecht University, 3508 TB Utrecht, The Netherlands
Methods: NMR, ESI-MS, FACE
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13. Compound ID: 939
Structure type: polymer chemical repeating unit
Trivial name: poly(glycosyl phosphate)
Compound class: CPS
Contained glycoepitopes: IEDB_136105,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_158539,IEDB_158556,IEDB_225177,IEDB_885813,IEDB_885823,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 279
Kaji E, Osa Y, Tanaike M, Hosokawa Y, Takayanagi H, Takada A "An alternative access to a trisaccharide repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 19A" -
Chemical and Pharmaceutical Bulletin 44 (1996) 437-440
A chemical synthesis has been achieved for β-D-ManNAc-(1→4)-α-D-Glc-(1→3)-L-Rha, a trisaccharide repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 19A, by stepwise link-up of the suitably functionalized, constituent sugar units. A beta-selective glycosylation of trimethylsilylethyl glucoside having free 4-OH with 2-(benzoyloxyimino)-2-deoxyglycosyl bromide, followed by manno-selective hydroboration, N-acetylation, and functionalization of the anomeric center (1-OSE→1-OH→1-F), gave a key disaccharide donor, β-D-ManNAc-(1→4)-α-D-Glc-(1→F. Ensuing glycosylation of an L-rhamnosyl acceptor with the donor substrate afforded, after deblocking, the target trisaccharide in 6.5% yield over 13 steps from D-glucose.
capsular polysaccharide, glycosylation, Streptococcus pneumoniae type 19A, 2-ulose oxime, β-D-mannosaminide, hydroboration
NCBI PubMed ID: 8998845Journal NLM ID: 0377775Publisher: Pharmaceutical Society Of Japan
Institutions: School of Pharmaceutical Sciences, Kitasato University, Japan, School of Pharmaceutical Sciences, Kitasato University
Methods: 13C NMR, 1H NMR, FAB-MS, TLC, chemical synthesis, chemical methods, UV, glycosylation
- Article ID: 332
Morona JK, Morona R, Paton JC "Comparative genetics of capsular polysaccharide biosynthesis in Streptococcus pneumoniae types belonging to serogroup 19" -
Journal of Bacteriology 181(17) (1999) 5355-5364
The genetic basis for the structural diversity of capsule polysaccharide (CPS) in Streptococcus pneumoniae serogroup 19 (consisting of types 19F, 19A, 19B, and 19C) has been determined for the first time. In this study, the genetic basis for the 19A and 19C serotypes is described, and the structures of all four serogroup 19 cps loci and their flanking sequences are compared. Transformation studies show that the structural difference between the 19A and 19F CPSs is likely to be a consequence of differences between their respective polysaccharide polymerase genes (cps19aI and cps19fI). The CPS of type 19C differs from that of type 19B by the addition of glucose. We have identified a single gene difference between the two cps loci (cps19cS), which is likely to encode a glucosyl transferase. The arrangement of the genes within the cps19 loci is highly conserved, with 13 genes (cps19A to -H and cps19K to -O) common to all four serogroup 19 members. These cps genes encode functions required for the synthesis of the shared trisaccharide component of the group 19 CPS repeat unit structures. Furthermore, the genetic differences between the group 19 cps loci identified are consistent with the CPS structures of the individual serotypes. Functions have been assigned to nearly all of the cps19 gene products, based on either gene complementation or similarity to other proteins with known functions, and putative biosynthetic pathways for production of all four group 19 CPSs have been proposed.
biosynthesis, genetic, Streptococcus, Streptococcus pneumoniae, capsular polysaccharide, type, serogroup
NCBI PubMed ID: 10464207Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: patonj@wch.sa.gov.au
Institutions: Molecular Microbiology Unit, Women's and Children's Hospital, North Adelaide, South Australia 5006, Department of Microbiology and Immunology, University of Adelaide, Adelaide, South Australia 5005
Methods: PCR, DNA sequencing, Southern blotting
- Article ID: 507
Pujar NS, Huang NF, Daniels CL, Dieter L, Gayton MG, Lee AL "Base hydrolysis of phosphodiester bonds in pneumococcal polysaccharides" -
Biopolymers 75(1) (2004) 71-84
A comprehensive study of the base hydrolysis of all phosphodiester bond-containing capsular polysaccharides of the 23-valent pneumococcal vaccine is described here. Capsular polysaccharides from serotypes 6B, 10A, 17F, 19A, 19F, and 20 contain a phosphodiester bond that connects the repeating units in these polysaccharides (also referred to as backbone phosphodiester bonds), and polysaccharides from serotypes 11A, 15B, 18C, and 23F contain a phosphodiester bond that links a side chain to their repeating units. Molecular weight measurements of the polysaccharides, using high performance size exclusion chromatography with tandem multiangle laser light scattering and refractive index detection, was used to evaluate the kinetics of hydrolysis. The measurement of molecular weight provides a high degree of sensitivity in the case of small extents of reaction, thus allowing reliable measurements of the kinetics over short times. Pseudo-first-order rate constants for these polysaccharides were estimated using a simple model that accounts for the polydispersity of the starting sample. It was found that the relative order of backbone phosphodiester bond instability due to base hydrolysis was 19A > 10A > 19F > 6B > 17F, 20. Degradation of side-chain phosphodiester bonds was not observed, although the high degree of sensitivity in measurements is lost in this case, due to the low contribution of the side chains to the total polysaccharide molecular weight. In comparison with literature data on pneumococcal polysaccharide 6A, 19A was found to be the more labile, and hence appears to be the most labile pneumococcal polysaccharide studied to date. The rate of hydrolysis increased at higher pH and in the presence of divalent cation, but the extent was lower than expected based on similar data on RNA. Finally, the differences in the phosphodiester bond stabilities were analyzed by considering stereochemical factors in these polysaccharides. These results also provide a framework for evaluation of molecular integrity of phosphodiester-bond-containing polysaccharides in different solution conditions. Copyright 2004 Wiley Periodicals, Inc. Biopolymers, 2004
base hydrolysis, phosphodiester bond, pneumococcal polysaccharide
NCBI PubMed ID: 15307199Journal NLM ID: 0372525Publisher: Wiley Interscience
Correspondence: hari_pujar@merck.com
Institutions: Merck Research Laboratories, Merck & Co., West Point, PA 19486
- Article ID: 1307
Zamze S, Martinez-Pomares L, Jones H, Taylor PR, Stillion RJ, Gordon S, Wong SY "Recognition of bacterial capsular polysaccharides and lipopolysaccharides by the macrophage mannose receptor" -
Journal of Biological Chemistry 277(44) (2002) 41613-41623
The in vitro binding of the macrophage mannose receptor to a range of different bacterial polysaccharides was investigated. The receptor was shown to bind to purified capsular polysaccharides from Streptococcus pneumoniae and to the lipopolysaccharides, but not capsular polysaccharides, from Klebsiella pneumoniae. Binding was Ca(2+)- dependent and inhibitable with d-mannose. A fusion protein of the mannose receptor containing carbohydrate recognition domains 4-7 and a full-length soluble form of the mannose receptor containing all domains external to the transmembrane region both displayed very similar binding specificities toward bacterial polysaccharides, suggesting that domains 4-7 are sufficient for recognition of these structures. Surprisingly, no direct correlation could be made between polysaccharide structure and binding to the mannose receptor, suggesting that polysaccharide conformation may play an important role in recognition. The full-length soluble form of the mannose receptor was able to bind simultaneously both polysaccharide via the carbohydrate recognition domains and sulfated oligosaccharide via the cysteine-rich domain. The possible involvement of the mannose receptor, either cell surface or soluble, in the innate and adaptive immune responses to bacterial polysaccharides is discussed
lipopolysaccharides, structure, Streptococcus pneumoniae, capsular polysaccharides, recognition, Klebsiella pneumoniae, Mannose, macrophage, receptor
NCBI PubMed ID: 12196537Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: susanne.zamze@jenner.ac.uk
Institutions: Edward Jenner Institute for Vaccine Research, Compton, Berkshire RG20 7NN, United Kingdom and the Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom
- Article ID: 1347
Abeygunawardana C, Williams TC, Sumner JS, Hennessey JP "Development and validation of an NMR-based identity assay for bacterial polysaccharides" -
Analytical Biochemistry 279(2) (2000) 226-240
A method utilizing NMR spectroscopy has been developed to confirm the identity of bacterial polysaccharides used to formulate a polyvalent pneumococcal polysaccharide vaccine. The method is based on 600 MHz proton NMR spectra of individual serotype-specific polysaccharides. A portion of the anomeric region of each spectrum (5.89 to 4.64 ppm) is compared to spectra generated for designated reference samples for each polysaccharide of interest. The selected region offers a spectral window that is unique to a given polysaccharide and is sensitive to any structural alteration of the repeating units. The similarity of any two spectral profiles is evaluated using a correlation coefficient (rho), where rho >/= 0.95 between a sample and reference profile indicates a positive identification of the sample polysaccharide. This method has been shown to be extremely selective in its ability to discriminate between serotype-specific polysaccharides, some of which differ by no more than a single glycosidic linkage. Furthermore, the method is rapid and does not require extensive sample manipulations or pretreatments. The method was validated as a qualitative identity assay and will be incorporated into routine quality control testing of polysaccharide powders to be used in preparation of the polyvalent pneumococcal vaccine PNEUMOVAX 23. The specificity and reproducibility of the NMR-based identity assay is superior to the currently used colorimetric assays and can be readily adapted for use with other bacterial polysaccharide preparations as well.
NMR, Bacterial, polysaccharide, Bacterial polysaccharide, polysaccharides, bacterial polysaccharides, assay, development, identity assay, method development, validation
NCBI PubMed ID: 10706792Publication DOI: 10.1006/abio.1999.447Journal NLM ID: 0370535Publisher: Academic Press
Correspondence: abey@merck.com
Institutions: Bioprocess and Bioanalytical Research, Merck Research Laboratories, West Point, Pensylvania, USA
Methods: NMR
- Article ID: 1556
Pujar NS, Huang NF, Daniels CL, Dieter L, Gayton MG, Lee AL "Erratum: Base hydrolysis of phosphodiester bonds in pneumococcal polysaccharides" -
Biopolymers 77(6) (2005) 378-379
No abstract
polysaccharide, Streptococcus, Research, hydrolysis, polysaccharides, phosphodiester, pneumococcal, PDF, P, pneumococcal polysaccharides
NCBI PubMed ID: 15761954Journal NLM ID: 0372525Publisher: Wiley Interscience
Institutions: WP17-301, P. O. Box 4, Merck Research Laboratories, Merck & Co., West Point, PA 19486
- Article ID: 3365
Nikolaev AV, Botvinko IV, Ross AJ "Natural phosphoglycans containing glycosyl phosphate units: structural diversity and chemical synthesis" -
Carbohydrate Research 342(3-4) (2007) 297-344
An anomeric phosphodiester linkage formed by a glycosyl phosphate unit and a hydroxyl group of another monosaccharide is found in many glycopolymers of the outer membrane in bacteria (e.g., capsular polysaccharides and lipopolysaccharides), yeasts and protozoa. The polymers (phosphoglycans) composed of glycosyl phosphate (or oligoglycosyl phosphate) repeating units could be chemically classified as poly(glycosyl phosphates). Their importance as immunologically active components of the cell wall and/or capsule of numerous microorganisms upholds the need to develop routes for the chemical preparation of these biopolymers. In this paper, we (1) present a review of the primary structures (known to date) of natural phosphoglycans from various sources, which contain glycosyl phosphate units, and (2) discuss different approaches and recent achievements in the synthesis of glycosyl phosphosaccharides and poly(glycosyl phosphates).
synthesis, structure, polysaccharides, Phosphoglycans, Anomeric phosphodiesters
NCBI PubMed ID: 17092493Publication DOI: 10.1016/j.carres.2006.10.006Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: a.v.nikolaev@dundee.ac.uk
Institutions: College of Life Sciences, Division of Biological Chemistry and Molecular Microbiology, University of Dundee, Dundee DD1 5EH, UK.
- Article ID: 3666
Bonaccorsi F, Catelani G, Oscarson S "A new route for the synthesis of Streptococcus pneumoniae 19F and 19A capsular polysaccharide fragments avoiding the b-mannosamine glycosylation step" -
Carbohydrate Research 344(12) (2009) 1442-1448
The recently described [Attolino, E.; Bonaccorsi, F.; Catelani, G.; D'Andrea, F. Carbohydr. Res. 2008, 343, 2545-2556.] β-D-MaNAcp-(1→4)-β-D-Glcp thiophenyl glycosyl donor 3 was used in alpha-glycosylation reactions of OH-2 and OH-3 of the suitably protected p-MeO-benzyl α-L-rhamnopyranoside acceptors 7 and 8. Glycosylation of the axial OH-2 of 7 took place in high yield (76%) and with acceptable stereoselectivity (alpha/beta=3.4) leading to the protected trisaccharide alpha-11, corresponding to the repeating unit of Streptococcus pneumoniae 19F. The same reaction on equatorial OH-3 of acceptor 8 gave the trisaccharide alpha-15, a constituent of the repeating unit of S. pneumoniae 19A, but in lower yield (41%) and without stereoselection (alpha/beta=1:1.3). Utilizing the introduced orthogonal protection of OH-1 and OH-4'', the trisaccharide alpha-11 was transformed into a trisaccharide building block suitable for the synthesis of its phosphorylated oligomers.
Streptococcus pneumoniae, glycoconjugate vaccines, oligosaccharide synthesis, glycosylation, Thioglycosides
NCBI PubMed ID: 19467536Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: G. Catelani
; S.Oscarson
Institutions: Dipartimento di Chimica Bioorganica e Biofarmacia, Universita di Pisa, Via Bonanno 33, I-56126 Pisa, Italy, Centre for Synthesis and Chemical Biology, UCD School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland
Methods: 13C NMR, 1H NMR, chemical synthesis
- Article ID: 3753
Park S, Parameswar AR, Demchenko AV, Nahm MH "Identification of a simple chemical structure associated with protective human antibodies against multiple pneumococcal serogroups" -
Infection and Immunity 77(8) (2009) 3374-3379
Streptococcus pneumoniae, a major human pathogen, expresses at least 91 serologically distinct carbohydrate capsules. Since pneumococcal vaccines are designed to elicit antibodies against many different capsular polysaccharides (PSs), it is important to identify the epitopes involved in eliciting anti-capsular PS antibodies. We investigated the epitopes recognized by Dob1, which is a hybridoma-secreting human immunoglobulin G2 antibody to the PS of serotype 6B (Y. Sun et al., Infect. Immun. 67:1172-1179, 1999). We found that Dob1 bound synthetic capsular carbohydrates Gal(1→3)α-D-Glcp(1→3)α-L-Rhap(1→3)Rib-ol and α-D-Glcp(1→3)α-L-Rhap(1→3)Rib-ol but did not bind α-L-Rhap(1→3)Rib-ol. The critical epitope α-D-Glcp(1→3)α-L-Rhap is found in the capsular PSs of serotypes 6A, 6B, 6C, and 19A but not in the 19F PS. Consistent with this observation, Dob1 bound to the PSs of serotypes 6A, 6B, 6C, and 19A but did not bind the 19F PS and 23 additional unrelated pneumococcal capsular PSs. Also, Dob1 could opsonize pneumococci expressing serotypes 6A, 6B, 6C, and 19A but did not opsonize 19F pneumococci. In addition, ca. 7% of immune sera (12 of 175 sera) had significant amounts of Dob1-like antibodies, i.e., reacted with 6B and 19A PSs, but not with 19F PS. Humans can produce antibodies to the Dob1 epitope and the antibodies to that epitope cross-react with the four serotypes 6A, 6B, 6C, and 19A that belong to different serogroups. This epitope may be useful for producing a totally synthetic, simple chemical structure that is capable of generating protective antibodies to multiple pneumococcal serogroups.
Bacterial, Streptococcus pneumoniae, antibodies, Bacterial Capsule, sepitopes, Pneumococcal Infections
NCBI PubMed ID: 19451241Publication DOI: 10.1128/IAI.00319-09Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: nahm@uab.edu
Institutions: University of Alabama at Birmingham, 35294-2170, USA
Methods: ELISA, serological methods, IEF
- Article ID: 4063
Lee CJ, Fraser BA, Boykins RA, Li JP "Effect of culture conditions on the structure of Streptococcus pneumoniae type 19A(57) capsular polysaccharide" -
Infection and Immunity 55(8) (1987) 1819-1823
The structural modifications and immunochemical activities of several Streptococcus pneumoniae type 19A polysaccharide (PS) preparations have been studied by sugar compositional analysis and immunodiffusion. The 19A PS preparations Lab-A-1 and Lab-A-3 and one PS isolated from 19A strain OB contained fucose (Fuc) and galactose (Gal) in addition to rhamnose (Rha) and glucose (Glc). In contrast, 19A PSs Lab-A-2 and Lab-B contained only Rha and Glc. Despite their different sugar compositions, these 19A preparations appeared to be identical in serologic activity as measured by immunodiffusion with rabbit 19A and 19F antisera. The 19A PS Lab-A-1 was separated into three fractions by DEAE-Sepharose CL-6B column chromatography with a NaCl gradient. Fraction II was the major peak with a yield of 72.9%. Fraction Ia contained Fuc and Gal, while fraction II contained Fuc, Gal, Rha, and Glc. Fractions Ia and Ib did not react with rabbit 19A antiserum. In contrast, 19A PS Lab-A-2 displayed only one peak, which was eluted by a NaCl gradient (0 to 0.6 M NaCl), and contained only Rha and Glc. The 19A PSs prepared from Lab-A and Centers for Disease Control (CDC) strains and grown in pneumococcal inoculum medium (PIM) and modified Holt medium were chromatographed on a DEAE-Sepharose CL-6B column, and the separated fractions were examined for their sugar composition. The fractions obtained from the 19A PSs Lab-A-PIM and CDC-PIM exhibited four sugar components, as observed for the PS Lab-A-1, while the separated fractions from the 19A PSs Lab-A-Holt and CDC-Holt displayed two sugar components, a pattern similar to that of PS Lab-A-2. Thus, the sugar compositions of 19A PS appeared to vary according to the type of culture medium used to grow the 19A organisms.
structure, Streptococcus pneumoniae, capsular polysaccharide, culture conditions
NCBI PubMed ID: 3610316Publication DOI: 10.1128/IAI.55.8.1819-1823.1987Journal NLM ID: 0246127Publisher: American Society for Microbiology
Institutions: Office of Biologics, Food and Drug Administration, Bethesda, MD, USA
Methods: sugar analysis, acid hydrolysis, serological methods, immunodiffusion assays
- Article ID: 4695
Kuttel M, Gordon M, Ravenscroft N "Comparative simulation of pneumococcal serogroup 19 polysaccharide repeating units with two carbohydrate force fields" -
Carbohydrate Research 390 (2014) 20-27
Streptococcus pneumoniae causes meningitis, pneumonia and severe invasive disease (IPD) in young children. Although widespread infant immunisation with the PCV7 seven-valent pneumococcal conjugate vaccine has led to a dramatic decrease in IPD, infections due to non-vaccine serotypes, particularly serotype 19A, have increased. As the 19F polysaccharide differs from 19A at a single linkage position, it was assumed that PCV7 (containing 19F) would cross-protect against 19A disease. However, vaccination with PCV7 results in only 26% effectiveness against IPD caused by 19A. We explored the conformations and dynamics of the polysaccharide repeating units from serotypes 19F and 19A, comparing free energy surfaces for glycosidic linkages with 100ns aqueous molecular dynamics simulations of the di- and trisaccharide components. All calculations were performed with both the CHARMM and the GLYCAM carbohydrate force fields to establish whether the choice of model affects the predicted molecular behaviour. Although we identified key differences between the force fields, overall they were in agreement in predicting a 19F repeating unit with a wider range of conformation families than the more restricted 19A trisaccharide. This suggests a probable conformational difference between the 19F and 19A polysaccharides, which may explain the low cross-protection of 19F vaccines against 19A disease.
molecular modelling, conformation, chemistry, disease, invasive, polysaccharide, serotype, Streptococcus, Streptococcus pneumoniae, repeating unit, trisaccharide, carbohydrate, molecular, Research, glycosidic linkage, infection, conformational, dynamics, molecular dynamics, serogroup, linkage, polysaccharides, families, position, surface, vaccines, calculation, computer, difference, Serotypes, component, aqueous, vaccine, pneumococcal, simulation, conjugate, children, pneumonia, conjugate vaccine, model, decrease, free, force field, meningitis, energy, vaccination, pneumococcal conjugate vaccine, Science, cross-protection, effectiveness, Infant
Publication DOI: 10.1016/j.carres.2014.02.026Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mkuttel@cs.uct.ac.za (M.Kuttel)
Institutions: Department of Computer Science, University of Cape Town, Cape Town, South Africa
Methods: conformation analysis, MD simulations, molecular modeling, MM3 force field
- Article ID: 4768
Kuttel MM, Jackson GE, Mafata M, Ravenscroft N "Capsular polysaccharide conformations in pneumococcal serotypes 19F and 19A" -
Carbohydrate Research 406 (2015) 27-33
Streptococcus pneumoniae is a significant pathogen in children. Although the PCV7 pneumococcal conjugate vaccine has reduced pneumococcal disease, non-vaccine serotype 19A infection has increased, despite expectations of cross-protection from vaccine serotype 19F. Serotype 19A is included in the new PCV13 vaccine, but not in PCV10. In the solution simulations of 19F and 19A oligosaccharide chains reported here, both polysaccharides form unstructured random coils, with inflexible repeat units linked by mobile phosphodiester linkages. However, there are clear conformational differences. In the 19F repeat unit, the rhamnose residue is nearly orthogonal to the other residues, whereas 19A has residues in similar orientations. This finding is corroborated by key inter-residue distances calculated from NMR NOESY experiments. Further, 19F is predominantly in extended conformations, whereas 19A exhibits a high prevalence of tight hairpin bends. These conformational differences may account for a lack of antibody cross-protection between serotypes 19F and 19A.
molecular modelling, Streptococcus pneumoniae, vaccine, pneumococcal, cross protection
NCBI PubMed ID: 25658063Publication DOI: 10.1016/j.carres.2014.12.013Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mkuttel@cs.uct.ac.za
Institutions: Department of Computer Science, University of Cape Town, Cape Town, South Africa
Methods: 13C NMR, 1H NMR, NMR-2D, conformation analysis, molecular modeling, MD simulation, polymer hydrodynamics
- Article ID: 4828
Geno KA, Gilbert GL, Song JY, Skovsted IC, Klugman KP, Jones C, Konradsen HB, Nahm MH "Pneumococcal Capsules and Their Types: Past, Present, and Future" -
Clinical Microbiology Reviews 28(3) (2015) 871-899
Streptococcus pneumoniae (the pneumococcus) is an important human pathogen. Its virulence is largely due to its polysaccharide capsule, which shields it from the host immune system, and because of this, the capsule has been extensively studied. Studies of the capsule led to the identification of DNA as the genetic material, identification of many different capsular serotypes, and identification of the serotype-specific nature of protection by adaptive immunity. Recent studies have led to the determination of capsular polysaccharide structures for many serotypes using advanced analytical technologies, complete elucidation of genetic basis for the capsular types, and the development of highly effective pneumococcal conjugate vaccines. Conjugate vaccine use has altered the serotype distribution by either serotype replacement or switching, and this has increased the need to serotype pneumococci. Due to great advances in molecular technologies and our understanding of the pneumococcal genome, molecular approaches have become powerful tools to predict pneumococcal serotypes. In addition, more-precise and -efficient serotyping methods that directly detect polysaccharide structures are emerging. These improvements in our capabilities will greatly enhance future investigations of pneumococcal epidemiology and diseases and the biology of colonization and innate immunity to pneumococcal capsules.
serotype, Streptococcus pneumoniae, vaccines, Pneumococcal Capsules
NCBI PubMed ID: 26085553Publication DOI: 10.1128/CMR.00024-15Journal NLM ID: 8807282Publisher: Washington, DC: American Society for Microbiology
Correspondence: Moon H. Nahm
Institutions: Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama, USA, Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology & Medical Research, Westmead Hospital, Wentworthville, New South Wales, Australia, Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Sydney, New South Wales, Australia, Division of Infectious Disease, Department of Internal Medicine, Korea University Guro Hospital, Seoul, South Korea, SSI Diagnostica, Division of Microbiology and Diagnostics, Statens Serum Institut, Copenhagen, Denmark, Pneumonia Program Strategy Team, Bill & Melinda Gates Foundation, Seattle, Washington, USA, Laboratory for Molecular Structure, NIBSC, South Mimms, Herts, United Kingdom, Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, USA
- Article ID: 4836
Berti F, Ravenscroft N "Characterization of Carbohydrate Vaccines by NMR Spectroscopy" -
Methods in Molecular Biology 1331 (2015) 189-209
Physicochemical techniques are a powerful tool for the structural characterization of carbohydrate-based vaccines. High-field Nuclear Magnetic Resonance (NMR) spectroscopy has been established as an extremely useful and robust method for tracking the industrial manufacturing process of these vaccines from polysaccharide bulk antigen through to the final formulation. Here, we describe the use of proton NMR for structural identity and conformity testing of carbohydrate-based vaccines.
carbohydrates, capsular polysaccharide, antigens, nuclear magnetic resonance spectroscopy, vaccines
NCBI PubMed ID: 26169742Publication DOI: 10.1007/978-1-4939-2874-3_12Journal NLM ID: 9214969Publisher: Springer
Correspondence: francesco.x.berti@gsk.com
Institutions: Research, GSK Vaccines, Via Fiorentina 1, 53100, Siena, Italy
- Article ID: 5440
Goyette-Desjardins G, Vinogradov E, Okura M, Takamatsu D, Gottschalk M, Segura M "Structure determination of Streptococcus suis serotypes 7 and 8 capsular polysaccharides and assignment of functions of the cps locus genes involved in their biosynthesis" -
Carbohydrate Research 473 (2019) 36-45
Streptococcus suis serotypes 7 and 8 are counted among the top six S. suis serotypes causing clinical disease in pigs. Yet, limited information is available on these serotypes. Since S. suis serotyping system is based upon capsular polysaccharide (CPS) antigenicity and the CPS is considered a major virulence factor for encapsulated pathogens, here we determined for the first time the chemical compositions and structures of serotypes 7 and 8 CPSs. Chemical and spectroscopic data gave the following repeating unit sequences: [3)L-Rha(α1-P-2)D-Gal(α1-4)D-GlcA(β1-3)D-FucNAc4N(α1-]n for serotype 7 and [2)L-Rha(α1-P-4)D-ManNAc(β1-4)D-Glc(α1-]n for serotype 8. As serotype 8 CPS is identical to Streptococcus pneumoniae type 19F CPS, dot-blot analyses showed a strong reaction of the 19F polysaccharide with reference anti-S. suis serotype 8 rabbit serum. A correlation between S. suis serotypes 7 and 8 sequences and genes of those serotypes' loci encoding putative glycosyltransferases and polymerases responsible for the biosynthesis of the repeating units was tentatively established. Knowledge of CPS structure and composition will contribute to better dissect the role of this bacterial component in the pathogenesis of the disease caused by S. suis serotypes 7 and 8.
capsular polysaccharide, carbohydrate structure, Serotype 8, Streptococcus suis, Serotype 7
NCBI PubMed ID: 30605786Publication DOI: 10.1016/j.carres.2018.12.009Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: M. Segura
Institutions: Swine and Poultry Infectious Diseases Research Center, Faculty of Veterinary Medicine, University of Montreal, 3200 Sicotte St., St-Hyacinthe, Quebec, J2S 2M2, Canada, Canadian Glycomics Network (GlycoNet), University of Alberta, 11227 Saskatchewan Dr., Edmonton, Alberta, T6G 2G2, Canada, National Research Council, 100 Sussex Dr., Ottawa, Ontario, K1A 0R6, Canada, Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan, The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu, Gifu, 501-1193, Japan
Methods: 13C NMR, 1H NMR, GLC-MS, gel filtration, NMR-2D, sugar analysis, 31P NMR, acid hydrolysis, Western blotting, bioinformatic analysis
- Article ID: 5473
Zou W, Li J, Vinogradov E, Cox A "Removal of cell wall polysaccharide in pneumococcal capsular polysaccharides by selective degradation via deamination" -
Carbohydrate Polymers 218 (2019) 199-207
Pneumococcal cell wall polysaccharide (C-PS), a contaminant in pneumococcal capsular polysaccharide (Pn-PS) vaccines is degraded by mild deamination of the 4-amino-2-acetamido-2,4,6-tri-deoxy-galactose (AAT) in C-PS, which was carried out by addition of 5% aqueous sodium nitrite to a solution of polysaccharide in 5% aqueous acetic acid. Glycosidic linkage and functional groups such as O-acetates, phosphodiesters, and pyruvates were preserved under the conditions. The small fragments from degraded C-PS were removed by ultrafiltration or dialysis to provide essentially C-PS free Pn-PS. Because of the presence of AAT in its structure the deamination is not suitable for the purification of type 1 Pn-PS. Meanwhile, the mass and NMR spectroscopic analysis on the deamination products suggests that both type 1 Pn-PS and C-PS degraded following a major pathway of 5,4-hydride shift, cleavage of AAT O5-C1 bond, C1 hemiacetal formation, and its hydrolysis to release neighboring GalA- in type 1 Pn-PS and GalNAc(6-O-PCho)- in C-PS
mechanism, degradation, deamination, cell wall polysaccharide, pneumococcal capsular polysaccharide
NCBI PubMed ID: 31221321Publication DOI: 10.1016/j.carbpol.2019.03.070Journal NLM ID: 8307156Publisher: Elsevier
Correspondence: W. Zou
Institutions: Human Health Therapeutic Research Center, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, K1A 0R6, Canada
Methods: 13C NMR, 1H NMR, gel filtration, sugar analysis, MS/MS, MS, dialysis, SEC-HPLC, ultrafiltration, mild deamination
- Article ID: 5791
Knirel YA, Van Calsteren M "Bacterial exopolysaccharides" -
Book: Comprehensive Glycoscience: From Chemistry to Systems Biology. Reference Module in Chemistry, Molecular Sciences and Chemical Engineering (2021) 1-75
Bacterial extracellular polysaccharides are known as a cell-bound capsule, a sheath, or a slime, which is excreted into the environment. They play an important role in virulence of medical bacteria and plant-to-symbiont interaction and are used for serotyping of bacteria and production of vaccines. Some exopolysaccharides have commercial applications in industry, and claims of health benefits have been documented for an increasing number of them. Exopolysaccharides have diverse composition and structure, and some contain sugar and non-sugar components that are found in bacterial carbohydrates only. The present article provides an updated collection of the data on exopolysaccharides of various classes of gram-negative and gram-positive bacteria reported until the end of 2019. When known, biosynthesis pathways of exopolysaccharides are treated in a summary manner. References are made to structure and biosynthesis relatedness between exopolysaccharides of different bacterial taxa as well as between bacterial polysaccharides and mammalian glycosaminoglycans.
polysaccharide structure, Gram-negative bacteria, capsule, Biofilm, polysaccharide biosynthesis, gram-positive bacteria, Monosaccharide composition, Bacterial exopolysaccharide, non-sugar component
Publication DOI: 10.1016/B978-0-12-819475-1.00005-5Publisher: Elsevier
Correspondence: marie-rose.vancalsteren@canada.ca; yknirel@gmail.com
Editors: Barchi J, Kamerling H
Institutions: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Saint-Hyacinthe Research and Development Centre, Agriculture and Agri-Food Canada, Saint-Hyacinthe, QC, Canada
- Article ID: 6073
Javed J, Mandal PK "Bacterial surface capsular polysaccharides from Streptococcus pneumoniae: A systematic review on structures, syntheses, and glycoconjugate vaccines" -
Carbohydrate Research 502 (2021) 108277
The polysaccharide capsule of Streptococcus pneumoniae constitutes the outermost surface structure of the organism and plays a critical role in virulence. The capsule is the target of current pneumococcal vaccines and glycoconjugates and has important medical and industrial applications. Widespread use of these vaccines is driving changes in serotype prevalence in disease. A massive array of sugars and glycosidic linkages experienced with complete diversity of potential polysaccharide structures. However, it is impossible to collect a sufficient quantity of glycan antigens for the preparation of CPS-based glycoconjugate vaccines from natural sources with high purity and for thorough biological evaluation. So nowadays, the development of a chemical synthetic strategy and their conjugation with a carrier protein to form synthetic glycoconjugate vaccines has been used to gain access on a large scale. This review provides a comprehensive summary of structures, synthesis as well as recent development of synthetic glycoconjugate vaccines, which will support research and may benefit the glycochemical and medical sciences.
synthesis, Streptococcus pneumoniae, glycoconjugate vaccines, polysaccharides, Pneumococcal Infections
NCBI PubMed ID: 33743443Publication DOI: 10.1016/j.carres.2021.108277Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: P.K. Mandal
Institutions: Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, BS-10/1, Sector 10, Jankipuram Extension, Sitapur Road, P.O. Box 173, Lucknow, 226 031, India, Academy of Scientific and Innovative Research (AcSIR), New Delhi, India
- Article ID: 6114
Morelli L, Lay L, Santana-Mederos D, Valdes-Balbin Y, Verez Bencomo V, van Diepen A, Hokke CH, Chiodo F, Compostella F "Glycan Array Evaluation of Synthetic Epitopes between the Capsular Polysaccharides from Streptococcus pneumoniae 19F and 19A" -
ACS Chemical Biology 16(9) (2021) 1671-1679
Vaccination represents the most effective way to prevent invasive pneumococcal diseases. The glycoconjugate vaccines licensed so far are obtained from capsular polysaccharides (CPSs) of the most virulent serotypes. Protection is largely limited to the specific vaccine serotypes, and the continuous need for broader coverage to control the outbreak of emerging serotypes is pushing the development of new vaccine candidates. Indeed, the development of efficacious vaccine formulation is complicated by the high number of bacterial serotypes with different CPSs. In this context, to simplify vaccine composition, we propose the design of new saccharide fragments containing chemical structures shared by different serotypes as cross-reactive and potentially cross-protective common antigens. In particular, we focused on Streptococcus pneumoniae (Sp) 19A and 19F. The CPS repeating units of Sp 19F and 19A are very similar and share a common structure, the disaccharide ManNAc-β-(1→4)-Glc (A-B). Herein, we describe the synthesis of a small library of compounds containing different combinations of the common 19F/19A disaccharide. The six new compounds were tested with a glycan array to evaluate their recognition by antibodies in reference group 19 antisera and factor reference antisera (reacting against 19F or 19A). The disaccharide A-B, phosphorylated at the upstream end, emerged as a hit from the glycan array screening because it is strongly recognized by the group 19 antisera and by the 19F and 19A factor antisera, with similar intensity compared with the CPSs used as controls. Our data give a strong indication that the phosphorylated disaccharide A-B can be considered a common epitope among different Sp 19 serotypes.
Streptococcus pneumoniae, capsular polysaccharides, glycan, disaccharide, glycoconjugate vaccine, vaccination, glycan epitopes
NCBI PubMed ID: 34469105Publication DOI: 10.1021/acschembio.1c00347Journal NLM ID: 101282906Publisher: Washington, DC: American Chemical Society
Correspondence: Fabrizio Chiodo < f.chiodo@amsterdamumc.nl>; Federica Compostella
Institutions: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Saldini 50, 20133 Milano, Italy, Department of Chemistry, University of Milan, Via Golgi 19, 20133 Milano, Italy, Finlay Vaccine Institute, 200 and 21 Street, 11600 Havana, Cuba, Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands, Italian National Research Council (CNR), Institute of Biomolecular Chemistry (ICB), Via Campi Flegrei 34, 80078 Pozzuoli, Italy
Methods: 13C NMR, 1H NMR, NMR-2D, TLC, 31P NMR, ESI-MS, chemical synthesis, chemical methods, immunization, glycan array analysis, immobilization
- Article ID: 6461
Shende N, Karale A, Bore P, Bhagade S, Gulhane A, Mallya AD, Dhere RM "Evaluation of structural modification induced activation of pneumococcal polysaccharide by GC-MS for the conjugate vaccine" -
Carbohydrate Research 531 (2023) 108878
Polysaccharide (Ps) activation evaluation is an imperative quality attribute in a conjugate vaccine. Pneumococcal polysaccharide (PnPs) serotypes 5, 6B, 14, 19A and 23F were cyanylated for 3 and 8 min. The cyanylated and non-cyanylated polysaccharides were methanolysed and derivatized to assess the activation of each sugar by GC-MS. The activation of 22 and 27% serotype 6B and 11 and 36% in serotype 23 F Ps at 3 and 8 min respectively showed controlled conjugation kinetics with CRM197 carrier protein estimated by SEC-HPLC and optimal absolute molar mass by SEC-MALS. The Glc and Gal are the most commonly activated sugars of all PnPs serotypes while N-acetyl sugars PneuNAc, GalNAc and Rha in serotypes 5, 14 and 19A respectively showed >50% activation which contributes to conjugate aggregate formation at 8 min compared to 3 min cyanylation. The GC-MS analysis of structural modifications at functional groups entails important information to characterize the activated polysaccharide for consistent conjugate vaccine manufacturing.
activation, conjugate vaccine, pneumococcal polysaccharide, GC-MS, cyanylation, SEC-HPLC, SEC-MALS
NCBI PubMed ID: 37390792Publication DOI: 10.1016/j.carres.2023.108878Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: A.D. Mallya
Institutions: Research and Development Department, Serum Institute of India Pvt. Ltd, Hadapsar, Pune, Maharashtra, 411028, India
Methods: GC-MS, methanolysis, SEC-MALS, conjugation, SEC-HPLC, CDAP-cyanylation, quantification
Expand this compound
Collapse this compound
14. Compound ID: 940
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_136105,IEDB_225177,IEDB_885813,IEDB_885823
The structure is contained in the following publication(s):
- Article ID: 280
Kaji E, Anabuki N, Zen S "Syntheses of three interglycosidic isomers of N-acetyl-b-D-mannosaminyl-L-rhamnoses associated with O-antigens of several gram-negative opportunistic pathogens" -
Chemical and Pharmaceutical Bulletin 43 (1995) 1441-1447
We achieved practical, highly stereoselective syntheses of three interglycosidic isomers of N-acetyl-β-D-mannosaminyl-L-rhamnoses, among which a β(1→4)-isomer corresponds to the repeating unit of the O-antigen of lipopolysaccharide (LPS) from the opportunistic pathogens Pseudomonas cepacia O5 and Pseudomonas aeruginosa X (Meitert). The other isomers are a β(1→2)-disaccharide, a constituent of LPS from Escherichia coli O1A, and an artificial β(1→3)-isomer. The disaccharides were obtained by simple three-step reaction sequences from 2-(benzoyloxyimino)-2-deoxyglycosyl halides (mannosamine progenitor). β-Selective glycosylations of appropriately protected L-rhamnosyl acceptors were performed. Subsequent reduction of the 2-acyloxyimino function to an amino group, N-acetylation, and removal of the protecting groups provided the target disaccharides. 13C NMR and nuclear Overhauser effect spectra proved to be useful for structural determination of the positional isomers of the disaccharides.
Lipopolysaccharide, O-antigen, Gram-negative bacteria, 2-amino-2-deoxy-D-mannose, β-3-D-mannosaminyl-L-rhamnose, 2-uiose oxime, opportunistic infection
NCBI PubMed ID: 7586068Journal NLM ID: 0377775Publisher: Pharmaceutical Society Of Japan
Institutions: School of Pharmaceutical Sciences, Kiiasaw University, Shirokane 5-9-1, Minato-ku, Tokyo IDS, Japan, School of Pharmaceutical Sciences, Kiiasaw University, Shirokane 5-9-1, Minato-ku, Tokyo IDS, Japan.
- Article ID: 405
Vinion-Dubiel AD, Goldberg JB "Lipopolysaccharide of Burkholderia cepacia complex" -
Journal of Endotoxin Research 9(4) (2003) 201-213
Burkholderia cepacia complex (Bcc) is a group of phenotypically similar, genetically distinct bacteria that are beneficial to the environment but can also cause severe human infections. Bcc are being exploited for use as bioremediation agents and as a way to combat agricultural plant diseases. However, Bcc can cause lung infections in patients with chronic granulomatous disease or cystic fibrosis often resulting in mortality of these patients. Since it is unclear what bacterial components are necessary for causing human infections, studies of Bcc have focused on identifying putative virulence factors. As in other Gram-negative bacteria, the lipopolysaccharide (LPS) of Bcc induces a strong immune response that can contribute to host cell damage. The unusual structure of Bcc LPS lowers the anionic charge of the Bcc cell surface, which inhibits the binding and subsequent effects of cationic antibiotics. These distinguishing features include the substitution of a Ko for a Kdo residue in the inner core oligosaccharide and Ara4N residues bound to phosphates of the lipid A backbone. The structures of O antigen subunits and the consequent serotypes will also be discussed, with particular reference to the O antigen biosynthetic loci of two Bcc strains.
Lipopolysaccharide, structure, Burkholderia, Burkholderia cepacia, O-antigen, complex, endotoxin, Re
NCBI PubMed ID: 12935351Publication DOI: 10.1177/09680519030090040101Journal NLM ID: 9433350Publisher: Maney Publishing
Institutions: Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville, VA 22908-0734, USA
- Article ID: 1311
Zähringer U, Rettenmaier H, Senchenkova SN, Knirel YA "Structure of a new 6-deoxy-alpha-D-talan from Burkholderia (Pseudomonas) plantarii strain DSM 6535, which is different from the O-chain of the lipopolysaccharide." -
Carbohydrate Research 300 (1997) 143-151
An O-acetylated homopolysaccharide of 6-deoxy-D-talose (6-deoxy-α-D-talan polymer) was isolated from Burkholderia_Pseudomonas plantarii strain DSM 6535 by extraction with 2-propanol. The structure (1) of the trisaccharide repeating unit of the polysaccharide was established by studies of the intact and O-deacetylated polysaccharides using methanolysis, methylation analysis, 1H and 13C NMR spectroscopy, including 2D COSY, heteronuclear 13C, 1H COSY, 1D NOE, and computer-assisted analysis of 1D 13C NMR spectra. The remaining material after extraction of the biomass with 2-propanol showed to be a lipopolysaccharide with an O-specific polysaccharide chain having a different structure (2), which has been found previously in lipopolysaccharides of a number of other Gram-negative bacteria. [formula: see text]
Lipopolysaccharide, structure, polysaccharide, 6-deoxy-D-talose, Burkholderia_Pseudomonas plantarii, 6-deoxy-D-tallose, Burkholderia Pseudomonas plantarii
NCBI PubMed ID: 9203339Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: uzaehr@fz-borstel.de
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Forschungszentrum Borstel, Zentrum fur Medizin und Biowissenschaften, D-23845 Borstel, Germany, BASF-AG, Main Laboratory, Ludwigshafen, Germany
Methods: methylation, NMR-2D, NMR, de-O-acetylation
- Article ID: 1453
Corsaro MM, De Castro C, Molinaro A, Parrilli M "Structure of lipopolysaccharides from phytopathogenic Gram-negative bacteria" -
Book: Recent Research Developments in Phytochemistry (2001) Vol. 5, 119-138
This review collects the structural data of lipopolysaccharide components arising from all phytopathogenic bacteria so far investigated. The structural approaches and the main biological role of these macromolecules are also reported.
Lipopolysaccharide, lipopolysaccharides, structure, core, lipid A, O-polysaccharide, gram negative bacteria
WWW link: https://books.google.ru/books/about/Recent_Research_Developments_in_Phytoche.html?id=5CJacgAACAAJ&redir_esc=yPublisher: Research Signpost, Trivandrum, India
Editors: Pandalai SG
Institutions: Dipartimento di Chimica Organica e Biochimica, Complesso Universitario Monte S.Angelo Via Cintia, 4, 80126 Napoli, Italy
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1837
Knirel YA, Kocharova NA, Shashkov AS, Varbanets LD, Kochetkov NK, Stanislavsky ES, Mashilova GM "Antigenic polysaccharides of bacteria. 17. Structure of O-specific polysaccharide chain of Pseudomonas aeruginosa X (Meitert) lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1268-1273
O-Specific polysaccharide composed of L-rhamnose and 2-acetamido-2-deoxy-D-mannose was obtained on mild acid degradation of P. aeruginosa X (Meitert classification) lipopolysaccharide. On the basis of non-destructive analis using 1H, 13C NMR spectroscopy and Klyne's rule calculation, as well as chemical methods (acid hydrolysis, methylation, Smith degradation), it was established that the polysaccharide is built up of disaccharide repeating units of the following structure: →4)-α-L-Rha-(1→3)-β-D-ManNAc-(1→.
NCBI PubMed ID: 2430585Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR
- Article ID: 2154
Cox AD, Wilkinson SG "Structures of the O-specific polymers from the lipopolysaccharides of the reference strains for Pseudomonas cepacia serogroups O3 and O5" -
Carbohydrate Research 195 (1989) 123-129
The putative O-specific polymers of lipopolysaccharides from two reference strains of Pseudomonas cepacia have been isolated and characterized. Both polymers have disaccharide repeating-units. Structure 1 was established for the O3 polymer, and structure 2 for the O5 polymer. Polymers with the same repeating units have been found previously as the O antigens of other bacteria. →2)-β-D-Ribf-(1→4)-α-D-GalpNAc-(1→ →4)-α-L-Rhap-(1→3)-β-D-ManpNAc-(1→.
NCBI PubMed ID: 2636041Publication DOI: 10.1016/0008-6215(89)85095-5Journal NLM ID: 0043535Publisher: Elsevier
Institutions: School of Chemistry, The University, Great Britain
Methods: 13C NMR, 1H NMR, methylation, GLC-MS, GLC, Smith degradation
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3321
Wang Z, Liu X, Dacanay A, Harrison BA, Fast M, Colquhoun DJ, Lund V, Brown LL, Li J, Altman E "Carbohydrate analysis and serological classification of typical and atypical isolates of Aeromonas salmonicida: a rationale for the lipopolysaccharide-based classification of A. salmonicida" -
Fish and Shellfish Immunology 23(5) (2007) 1095-1106
The cell envelope of Aeromonas salmonicida contains a lipopolysaccharide (LPS) essential for the physical integrity and functioning of bacterial cell membrane. Using a recently developed in-source fragmentation technique, we screened 39 typical and atypical isolates of A. salmonicida and established their O-chain polysaccharide structure by capillary electrophoresis-mass spectrometry (CE-MS), compositional and linkage analyses and comparison to the previously determined O-chain polysaccharide structure of A. salmonicida strain A449. These studies have demonstrated that A. salmonicida isolates fall into three distinct structural types, types A-C, based on chemical structures of their respective O-chain polysaccharide components. Subsequent immunoblotting and serological studies with salmon polyclonal antisera produced to formalin-fixed cells of A. salmonicida strains A449, N4705 and 33659 representing three structural types A-C revealed that variations in the O-chain polysaccharide structure have led to significant serological differences between strains belonging to type A and non-type A, where non-type A species include chemically separated structural types B and C. Due to the presence of common antigenic determinants shared by their respective O-chain polysaccharide components, serological cross-reactions were observed between A. salmonicida strains belonging to structural types B and C. These findings suggest the possibility of developing LPS-based classification system of A. salmonicida sub-species consisting of two serologically distinct types, type A and non-type A
Lipopolysaccharide, mass spectrometry, classification, Aeromonas salmonicida
NCBI PubMed ID: 17658271Journal NLM ID: 9505220Publisher: Academic Press
Correspondence: eleonora.altman@nrc-cnrc.gc.ca
Institutions: Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, ON, K1A 0R6, Canada
Methods: SDS-PAGE, serological methods, analytical methods, immunoblotting, CE-MS
- Article ID: 3812
Altman E, Li J "Characterization of polysaccharides using mass spectrometry for bacterial serotyping" -
Book: Methods in Molecular Biology (2010) Vol. 600, 245-257
Mass spectrometry provides a rapid and reliable method for characterization of bacterial polysaccharides. Application of the in-source fragmentation technique to promote the formation of structurally relevant repeating units of heterogeneous capsular polysaccharides and O-chain polysaccharides has proven to be particularly useful for detection of non-carbohydrate functionalities and subtle differences arising across bacterial serotypes. Here we discuss application of these methods to the direct analysis of bacterial cells allowing for rapid analysis of cell surface polysaccharide antigens and providing a basis for serological typing and epidemiological surveillance studies of human and animal pathogens.
Lipopolysaccharide, mass spectrometry, classification, serotyping, preconcentration
NCBI PubMed ID: 19882133Publisher: Totowa, NJ: Humana Press
Editors: Holst O, Walker JM, Beck A
Institutions: Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada
Methods: methylation, GC-MS, acid hydrolysis, composition analysis, methanolysis, CE-MS
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4329
Knirel YA "Structure of O-antigens" -
Book: Bacterial lipopolysaccharides: Structure, chemical synthesis, biogenesis and interaction with host cells (2011) Chapter 3, 41-115
The lipopolysaccharide (LPS) is the major constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. Its lipid A moiety is embedded in the membrane and serves as an anchor for the rest of the LPS molecule. The outermost repetitive glycan region of the LPS is linked to the lipid A through a core oligosaccharide (OS), and is designated as the O-specific polysaccharide (O-polysaccharide, OPS) or O-antigen. The O-antigen is the most variable portion of the LPS and provides serological specificity, which is used for bacterial serotyping. The OPS also provides protection to the microorganisms from host defenses such as complement mediated killing and phagocytosis, and is involved in interactions of bacteria with plants and bacteriophages. Studies of the OPSs ranging from the elucidation of their chemical structures and conformations to their biological and physico-chemical properties help improving classification schemes of Gram-negative bacteria. Furthermore, these studies contributed to a better understanding of the mechanisms of pathogenesis of infectious diseases, as well as provided information to develop novel vaccines and diagnostic reagents.
Lipopolysaccharide, synthesis, lipopolysaccharides, structure, Bacterial, host, O-antigen, O antigen, cell, O antigens, O-antigens, chemical, interaction, cells, PDF, chemical synthesis, biogenesis
Publication DOI: 10.1007/978-3-7091-0733-1_3Publisher: Springer
Correspondence: knirel@ioc.ac.ru
Editors: Knirel YA, Valvano MA
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 5143
Cloutier M, Muru K, Ravicoularamin G, Gauthier C "Polysaccharides from Burkholderia species as targets for vaccine development, immunomodulation and chemical synthesis" -
Natural Product Reports 35(12) (2018) 1251-1293
Burkholderia species are a vast group of human pathogenic, phytopathogenic, and plant- or environment-associated bacteria. B. pseudomallei, B. mallei, and B. cepacia complex are the causative agents of melioidosis, glanders, and cystic fibrosis-related infections, respectively, which are fatal diseases in humans and animals. Due to their high resistance to antibiotics, high mortality rates, and increased infectivity via the respiratory tract, B. pseudomallei and B. mallei have been listed as potential bioterrorism agents by the Centers for Disease Control and Prevention. Burkholderia species are able to produce a large network of surface-exposed polysaccharides, i.e., lipopolysaccharides, capsular polysaccharides, and exopolysaccharides, which are virulence factors, immunomodulators, major biofilm components, and protective antigens, and have crucial implications in the pathogenicity of Burkholderia-associated diseases. This review provides a comprehensive and up-to-date account regarding the structural elucidation and biological activities of surface polysaccharides produced by Burkholderia species. The chemical synthesis of oligosaccharides mimicking Burkholderia polysaccharides is described in detail. Emphasis is placed on the recent research efforts toward the development of glycoconjugate vaccines against melioidosis and glanders based on synthetic or native Burkholderia oligo/polysaccharides.
lipopolysaccharides, Burkholderia, capsular polysaccharides, Oligosaccharides, glycoconjugate vaccines, antigens, exopolysaccharides, surface polysaccharide, virulence factor, Biofilm, chemical synthesis, bioterrorism
Publication DOI: 10.1039/C8NP00046HJournal NLM ID: 8502408Publisher: London: Royal Society of Chemistry
Correspondence: charles.gauthier@iaf.inrs.ca
Institutions: INRS-Institut Armand-Frappier, Universite du Quebec, 531, boul. des Prairies, Laval, Canada
- Article ID: 5488
Perepelov AV, Li X, Xu C, Filatov AV, Shashkov AS, Senchenkova SN, Liu B "Structure elucidation and gene cluster characterization of the O-antigen of Vibrio cholerae O14" -
Carbohydrate Research 474 (2019) 67-71
The O-polysaccharide (O-antigen) of Vibrio cholerae O14 was studied using chemical analyses and 1D and 2D NMR spectroscopy. The following structure of the repeating unit of the O-antigen was established: where GlcpN(SHb) indicates 2-deoxy-2-[(S)-3-hydroxybutanoylamino]-d-glucose. We found that Vibrio cholerae O14 is similar to that of O-polysaccharide of Azospirillum brasilense S17, which has been reported earlier. Moreover, we predicted functions of all the genes in the O-antigen gene cluster according to the structure established. Our study enriches the existing O-antigen database of Vibrio cholerae, and further facilitates the bacterial serotype identification.
Lipopolysaccharide, O-antigen, O-specific polysaccharide, bacterial polysaccharide structure, Vibrio cholerae, O-antigen gene cluster, GRASS
NCBI PubMed ID: 30763794Publication DOI: 10.1016/j.carres.2019.01.007Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: A.V. Perepelov
Institutions: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, TEDA Institure of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, China
Methods: 13C NMR, 1H NMR, NMR-2D, DNA sequencing, sugar analysis, acid hydrolysis, GLC, Smith degradation, GPC, delipidation, function analysis of gene clusters, function analysis of gene cluster, 13C NMR analysis by GRASS
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15. Compound ID: 945
Structure type: polymer chemical repeating unit
Compound class: CPS, O-polysaccharide, K-antigen, O-antigen
Contained glycoepitopes: IEDB_885813
The structure is contained in the following publication(s):
- Article ID: 285
Keenleyside WJ, Whitfield C "A novel pathway for O-polysaccharide biosynthesis in Salmonella enterica serovar Borreze" -
Journal of Biological Chemistry 271 (1996) 28581-28592
The plasmid-encoded gene cluster for O:54 O-polysaccharide synthesis in Salmonella enterica serovar Borreze (rfbO:54) contains three genes that direct synthesis of a ManNAc homopolymer with alternating β1,3 and β1,4 linkages. In Escherichia coli K-12, RfbAO:54 adds the first ManNAc residue to the Rfe (UDP-GlcpNAc::undecaprenylphosphate GlcpNAc-1-phosphate transferase)- modified lipopolysaccharide core. Hydrophobic cluster analysis of RfbAO:54 indicates this protein belongs to the ExoU family of nonprocessive β-glycosyltransferases. Two putative catalytic residues and a potential substrate-binding motif were identified in RfbAO:54. Topological analysis of RfbBO:54 predicts four transmembrane domains and a large central cytoplasmic domain. The latter shares homology with a similar domain in the processive β-glycosyltransferases Cps3S of Streptococcus pneumoniae and HasA of Streptococcus pyogenes. Hydrophobic cluster analysis of RfbBO:54 and Cps3S indicates both possess the structural features characteristic of the HasA family of processive β-glycosyltransferases. Four potential catalytic residues and a putative substrate-binding motif were identified in RfbBO:54. In ∆rfb E. coli K-12, RfbAO:54 and RfbBO:54 direct synthesis of smooth O:54 lipopolysaccharide, indicating that this O-polysaccharide involves a novel pathway for O-antigen transport. Based on sequence and structural conservation, 15 new ExoU-related and 17 new HasA-related transferases were identified.
Lipopolysaccharide, biosynthesis, O-antigen, O-polysaccharide, Salmonella, Salmonella enterica
NCBI PubMed ID: 8910488Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: cwhitfie@micro.uoguelph.ca
Institutions: From the Department of Microbiology, University of Guelph, Guelph, Ontario N1G2W1, Canada.
- Article ID: 286
Keenleyside WJ, Whitfield C "Lateral transfer of rfb genes: a mobilizable ColE1-type plasmid carries the rfbO:54 (O:54 antigen biosynthesis) gene cluster from Salmonella enterica serovar Borreze" -
Journal of Bacteriology 177(18) (1995) 5247-5253
Plasmid pWQ799 is a 6.9-kb plasmid isolated from Salmonella enterica serovar Borreze. Our previous studies have shown that the plasmid contains a functional biosynthetic gene cluster for the expression of the O:54 lipopolysaccharide O-antigen of this serovar. The minimal replicon functions of pWQ799 have been defined, and a comparison with nucleotide and protein databases revealed this replicon to be virtually identical to ColEl. This is the first report of involvement of ColEl-related plasmids in O-antigen expression. The replicon of pWQ799 is predicted to encode two RNA molecules, typical of other ColEl-type plasmids. RNAII, the putative replication primer from pWQ799, shares regions of homology with RNAII from ColEl. RNAI is an antisense regulator of DNA replication in ColEl-related plasmids. The coding region for RNAI from pWQ799 shares no homology with any other known RNAI sequence but is predicted to adopt a secondary structure characteristic of RNAI molecules. pWQ799 may therefore represent a new incompatibility group within this family. pWQ799 also possesses cer, rom, and mob determinants, and these differ minimally from those of ColEl. The plasmid is mobilizable in the presence of either the broad-host-range helper plasmid pRK2013 or the Inclj plasmid R64drd86. Mobilization and transfer of pWQ799 to other organisms provides the first defined mechanism for lateral transfer of O-antigen biosynthesis genes in S. enterica and explains both the distribution of related plasmids and coexpression of the O:54 factor with other O-factors in different Salmonella serovars. The base composition of the pWQ799 replicon sequences gives an average percent G+C value typical of Salmonella spp. In contrast, the percent G+C value is dramatically lower within rfb0:54, consistent with the possibility that the cluster was acquired from an organism with a much lower G+C composition.
biosynthesis, transfer, antigen, O-antigen, rfb, gene cluster, Salmonella, plasmid, Salmonella enterica, lateral
NCBI PubMed ID: 7545154Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: cwhitfie@micro.uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, Canada NIG 2W1
- Article ID: 739
Keenleyside WJ, Clarke AJ, Whitfield C "Identification of residues involved in catalytic activity of the inverting glycosyl transferase WbbE from Salmonella enterica serovar Borreze" -
Journal of Bacteriology 183(1) (2001) 77-85
Synthesis of the O:54 O antigen of Salmonella enterica is initiated by the nonprocessive glycosyl transferase WbbE, assigned to family 2 of the glycosyl transferase enzymes (GT2). GT2 enzymes possess a characteristic N-terminal domain, domain A. Based on structural data from the GT2 representative SpsA (S. J. Charnock and G. J. Davies, Biochemistry 38:6380-6385, 1999), this domain is responsible for nucleotide binding. It possesses two invariant Asp residues, the first forming a hydrogen bond to uracil and the second coordinating a Mn(2+) ion. Site-directed replacement of Asp41 (D41A) of WbbE, the analogue of the first Asp residue of SpsA, revealed that this is not required for activity. WbbE possesses three Asp residues near the position analogous to the second conserved residue. Whereas D95A reduced WbbE activity, activity in D93A and D96A mutants was abrogated, suggesting that either D93 or D96 may coordinate the Mn(2+) ion. Our studies also identified a C-terminal region of sequence conservation in 22 GT2 members, including WbbE. SpsA was not among these. This region is characterized by an ED(Y) motif. The Glu and Asp residues of this motif were individually replaced in WbbE. E180D in WbbE had greatly reduced activity, and an E180Q replacement completely abrogated activity; however, D181E had no effect. E180 is predicted to reside on a turn. Combined with the alignment of the motif with potential catalytic residues in the GT2 enzymes ExoM and SpsA, we speculate that E180 is the catalytic residue of WbbE. Sequence and predicted structural divergence in the catalytic region of GT2 members suggests that this is not a homogeneous family
synthesis, antigen, chemistry, genetics, metabolism, microbiology, potential, Sequence Analysis, DNA, structural, Support, Non-U.S.Gov't, O-antigen, O antigen, hydrogen, hydrogen bond, ion, transferase, conserved, Molecular Sequence Data, mutant, mutants, glycosyltransferases, Salmonella, conservation, activity, identification, motif, region, families, position, reduced, biochemistry, Salmonella enterica, sequence, structure-activity relationship, enzyme, binding, domain, Enzymes, glycosyl, Cell Membrane, mutagenesis, effect, protein structure, aspartic acid, analogue, enzymology, amino acid sequence, amino acid motifs, catalysis, Catalytic Domain, Consensus Sequence, divergence, Glucosyltransferases, Site-Directed, Secondary, Sequence Alignment, United States
NCBI PubMed ID: 11114903Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: cwhitfie@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada
- Article ID: 2393
van der Kaaden A, Gerwig GJ, Kamerling JP, Vliegenthart JFG, Tiesjema RH "Structure of the capsular antigen of Neisseria meningitidis serogroup K" -
European Journal of Biochemistry 152 (1985) 663-668
The capsular antigen isolated from the culture liquid of a Neisseria meningitidis serogroup K(1 811) fermentation consists of the 2-acetamido-2-deoxymannuronic acid disaccharide repeating unit as follows: (formula in text). The polysaccharide is 0-acetylated at the non-giycosylated C-4. Structural evidence has been obtained from sugar analysis, methylaion analysis, as well a 1H and 13C NMR spectroscopy.
NCBI PubMed ID: 3932071Publication DOI: 10.1111/j.1432-1033.1985.tb09246.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Bio-Organic Chemistry, State University of Utrecht, and National Institute for Public Health and Environmental Hygiene, Bilthoven
Methods: 13C NMR, 1H NMR
- Article ID: 2779
Keenleyside WJ, Perry M, MacLean L, Poppe C, Whitfield C "A plasmid-encoded rfbO:54 gene cluster is required for biosynthesis of the O:54 antigen in Salmonella enterica serovar Borreze" -
Molecular Microbiology 11 (1994) 437-448
Previous studies demonstrated that the presence of a 7-8 kb plasmid is correlated with expression of the lipopolysaccharide (LPS) O:54 antigen in several Salmonella enterica serovars. In this study, a 6.7 kb plasmid from a field isolate of S. enterica serovar Borreze was shown to encode enzymes responsible for the synthesis of the O:54 polysaccharide. Curing the plasmid results in simultaneous loss of smooth O-polysaccharide-substituted LPS molecules and O:54 serotype. SDS-PAGE analysis of other O:54 isolates indicated that the O:54 O-polysaccharide can be co-expressed with an additional O-polysaccharide, likely encoded by chromosomal genes. The structure of the O:54 polysaccharide was determined by a combination of chemical and nuclear magnetic resonance (NMR) methods and was found to be an unusual homopolymer of N-acetylmannosamine (D-ManNAc) residues. The polysaccharide contained a disaccharide repeating unit with the structure: →4)-β-D-ManpNAc-(1→3)-β-D-ManpNAc-(1→. This structure does not resemble other O-polysaccharides in S. enterica. To examine the role played by plasmid functions in synthesis of the O:54 polysaccharide, the 6.7 kb plasmid was cloned to produce a hybrid plasmid (pWQ800) in pGEM-7Zf(+). In Escherichia coli K-12 ∆rfb, pWQ800 directed the synthesis of authentic O:54 polysaccharide. Polymerized O:54 polysaccharide was also produced in S. enterica serovar Typhimurium rfb and rfc mutants. From these data, we conclude that pWQ800 carries the rfbO:54 gene cluster and synthesis of the O:54 polysaccharides does not require host chromosomal rfb functions. However, synthesis of the O:54 polysaccharide requires the function of the rfe and rffE genes which are part of the gene cluster encoding enzymes involved in biosynthesis of enterobacterial common antigen. The rffE gene product synthesizes the O:54 precursor, uridine diphospho-N-acetylmannosamine. This is the first description of a plasmid-encoded rfb gene cluster in Salmonella.
NCBI PubMed ID: 7512186Journal NLM ID: 8712028Publisher: Blackwell Publishing
Institutions: Department of Microbiology, University of Guelph, Ontario, Canada
Methods: 13C NMR
- Article ID: 3847
Gajdus J, Glosnicka R, Szafranek J "Primary structure of Salmonella spp. O-antigens" -
Wiadomosci Chemiczne [Polish] 60(9-10) (2006) 621-653
Salmonella spp. are pathogenic Gram-negative bacteria that belong to Enterobacteriaceae family with lipopolysaccharide (LPS) as a constituent of cell wall. This is an integral component of the outer membrane of the wall. Salmonella smooth (S) forms produce LPS, which is composed of three parts, chemically bonded together viz. polysaccharide O-antigen, oligosaccharide core region and lipid A. Antigens O (O-PS) together with H flagella antigens are the foundation of serological classification of these bacteria. O-chain, which is built with up to 50 oligosaccharide repeating units, is one of the products of mild acidic hydrolysis of LPS. Due to the fact that polysaccharide antigens are the sites of specific antibody complexing, any difference in primary and secondary structures of O-antigens reflect serological specificity of bacteria. Taking this fact into consideration, we can distinguish about 2541 Salmonella serotypes with O and H antigenic formulas defined [4]. In this review we present 55 chemical structures of O-antigenic repeating units of Salmonella strains including their heterogeneity structures. The structures can have 22 different monosaccharide residues usually in 3 to 6 sugar repeating units. We describe here selected chemical and spectroscopic (MS, NMR) methods for primary structure examination of these bacterial O-PS. Enzymatic and immunochemical methods are also described. Cross-reactions of Salmonella spp. with any other bacteria or blood group A, B, 0 antigens are explained on the molecular level. Thus, structural assignments of somatic antigens of Salmonella spp. allow us to understand the molecular level of the classification system of these bacteria.
NMR spectroscopy, O-antigens, Salmonella, MS, primary structure
WWW link: http://baztech.icm.edu.pl/baztech/cgi-bin/btgetdoc.cgi?BUS2-0016-0014Publisher: Polish Chemical Society
Correspondence: jerzyg@chemik.chem.univ.gda.pl
Institutions: Wydzial Chemii, Uniwersytet Gdanski, ul. Sobieskiego 18, 80-952 Gdansk
- Article ID: 4329
Knirel YA "Structure of O-antigens" -
Book: Bacterial lipopolysaccharides: Structure, chemical synthesis, biogenesis and interaction with host cells (2011) Chapter 3, 41-115
The lipopolysaccharide (LPS) is the major constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. Its lipid A moiety is embedded in the membrane and serves as an anchor for the rest of the LPS molecule. The outermost repetitive glycan region of the LPS is linked to the lipid A through a core oligosaccharide (OS), and is designated as the O-specific polysaccharide (O-polysaccharide, OPS) or O-antigen. The O-antigen is the most variable portion of the LPS and provides serological specificity, which is used for bacterial serotyping. The OPS also provides protection to the microorganisms from host defenses such as complement mediated killing and phagocytosis, and is involved in interactions of bacteria with plants and bacteriophages. Studies of the OPSs ranging from the elucidation of their chemical structures and conformations to their biological and physico-chemical properties help improving classification schemes of Gram-negative bacteria. Furthermore, these studies contributed to a better understanding of the mechanisms of pathogenesis of infectious diseases, as well as provided information to develop novel vaccines and diagnostic reagents.
Lipopolysaccharide, synthesis, lipopolysaccharides, structure, Bacterial, host, O-antigen, O antigen, cell, O antigens, O-antigens, chemical, interaction, cells, PDF, chemical synthesis, biogenesis
Publication DOI: 10.1007/978-3-7091-0733-1_3Publisher: Springer
Correspondence: knirel@ioc.ac.ru
Editors: Knirel YA, Valvano MA
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 5866
Wear SS, Hunt BA, Clarke BR, Whitfield C "Analysis of the Topology and Active-Site Residues of WbbF, a Putative O-Polysaccharide Synthase from Salmonella enterica Serovar Borreze" -
Journal of Bacteriology 202(5) (2020) e00625-19
Bacterial lipopolysaccharides are major components and contributors to the integrity of Gram-negative outer membranes. The more conserved lipid A-core part of this complex glycolipid is synthesized separately from the hypervariable O-antigenic polysaccharide (OPS) part, and they are joined in the periplasm prior to translocation to the outer membrane. Three different biosynthesis strategies are recognized for OPS biosynthesis, and one, the synthase-dependent pathway, is currently confined to a single example: the O:54 antigen from Salmonella enterica serovar Borreze. Synthases are complex enzymes that have the capacity to both polymerize and export bacterial polysaccharides. Although synthases like cellulose synthase are widespread, they typically polymerize a glycan without employing a lipid-linked intermediate, unlike the O:54 synthase (WbbF), which produces an undecaprenol diphosphate-linked product. This raises questions about the overall similarity between WbbF and conventional synthases. In this study, we examine the topology of WbbF, revealing four membrane-spanning helices, compared to the eight in cellulose synthase. Molecular modeling of the glycosyltransferase domain of WbbF indicates a similar architecture, and site-directed mutagenesis confirmed that residues important for catalysis and processivity in cellulose synthase are conserved in WbbF and required for its activity. These findings indicate that the glycosyltransferase mechanism of WbbF and classic synthases are likely conserved despite the use of a lipid acceptor for chain extension by WbbF.IMPORTANCE Glycosyltransferases play a critical role in the synthesis of a wide variety of bacterial polysaccharides. These include O-antigenic polysaccharides, which form the distal component of lipopolysaccharides and provide a protective barrier important for survival and host-pathogen interactions. Synthases are a subset of glycosyltransferases capable of coupled synthesis and export of glycans. Currently, the O:54 antigen of Salmonella enterica serovar Borreze involves the only example of an O-polysaccharide synthase, and its generation of a lipid-linked product differentiates it from classical synthases. Here, we explore features conserved in the O:54 enzyme and classical synthases to shed light on the structure and function of the unusual O:54 enzyme.
Lipopolysaccharide, O antigens, glycosyltransferase, synthase, polysaccharide biosynthesis, polysaccharide export
NCBI PubMed ID: 31792013Publication DOI: 10.1128/JB.00625-19Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: cwhitfie@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada
Methods: DNA techniques, genetic methods, biochemical methods, immunofluorescence microscopy
- Article ID: 5869
Whitfield C, Williams DM, Kelly SD "Lipopolysaccharide O-antigens-bacterial glycans made to measure" -
Journal of Biological Chemistry 295(31) (2020) 10593-10609
Lipopolysaccharides are critical components of bacterial outer membranes. The more conserved lipid A part of the lipopolysaccharide molecule is a major element in the permeability barrier imposed by the outer membrane and offers a pathogen-associated molecular pattern recognized by innate immune systems. In contrast, the long-chain O-antigen polysaccharide (O-PS) shows remarkable structural diversity and fulfills a range of functions, depending on bacterial lifestyles. O-PS production is vital for the success of clinically important Gram-negative pathogens. The biological properties and functions of O-PSs are mostly independent of specific structures, but the size distribution of O-PS chains is particularly important in many contexts. Despite the vast O-PS chemical diversity, most are produced in bacterial cells by two assembly strategies, and the different mechanisms employed in these pathways to regulate chain-length distribution are emerging. Here, we review our current understanding of the mechanisms involved in regulating O-PS chain-length distribution and discuss their impact on microbial cell biology.
polysaccharide, O-antigen, Gram-negative bacteria, glycosyltransferase, outer membrane, lipopolysaccharide (LPS), glycan biosynthesis, bacterial pathogenesis, antigenic diversity, cell surface
NCBI PubMed ID: 32424042Publication DOI: 10.1074/jbc.REV120.009402Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: cwhitfie@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada
- Article ID: 5871
Yakovlieva L, Walvoort MTC "Processivity in Bacterial Glycosyltransferases" -
ACS Chemical Biology 15(1) (2020) 3-16
Extracellular polysaccharides and glycoproteins of pathogenic bacteria assist in adherence, autoaggregation, biofilm formation, and host immune system evasion. As a result, considerable research in the field of glycobiology is dedicated to study the composition and function of glycans associated with virulence, as well as the enzymes involved in their biosynthesis with the aim to identify novel antibiotic targets. Especially, insights into the enzyme mechanism, substrate binding, and transition-state structures are valuable as a starting point for rational inhibitor design. An intriguing aspect of enzymes that generate or process polysaccharides and glycoproteins is the level of processivity. The existence of enzymatic processivity reflects the need for regulation of the final glycan/glycoprotein length and structure, depending on the role they perform. In this Review, we describe the currently reported examples of various processive enzymes involved in polymerization and transfer of sugar moieties, predominantly in bacterial pathogens, with a focus on the biochemical methods, to showcase the importance of studying processivity for understanding the mechanism.
processivity, Apparent processivity, Distributivity, Hyperglycosylation, Intrinsic processivity, Semiprocessive
NCBI PubMed ID: 31750644Publication DOI: 10.1021/acschembio.9b00619Journal NLM ID: 101282906Publisher: Washington, DC: American Chemical Society
Correspondence: m.t.c.walvoort@rug.nl
Institutions: Stratingh Institute for Chemistry, University of Groningen, 9747 AG Groningen, The Netherlands
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