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1. Compound ID: 1056
Structure type: polymer chemical repeating unit
Trivial name: repeating unit, type Y polysaccharide, oligosaccharide repeating unit
Compound class: O-polysaccharide, O-antigen, CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 300
Lee CH, Frasch CE "Quantification of bacterial polysaccharides by the purpald assay: Measurement of periodate-generated formaldehyde from glycol in the repeating unit" -
Analytical Biochemistry 296(1) (2001) 73-82
We have adapted the purpald assay for measurement of bacterial polysaccharides (PS) containing substituted and/or unsubstituted glycol (SG or UG) in residues such as glycerol, ribitol, arabinitol, furanosyl galactose, and sialyl. For the purpald assay of UG-containing PS, 50 microL of PS samples was consecutively reacted with 50 microL of 16 mM NaIO4 for 20 min, 50 microL of 136 mM purpald reagent in 2 N NaOH for 20 min, and 50 microL of 64 mM NaIO4 for 20 min in a 96-well tissue culture plate followed by a measurement of absorbance at 550 nm with a plate reader. For SG-containing PS, conversion of SG to UG with 25 micro;L of 0.3 N NaOH, 1 h at room temperature for de-O-acetylation followed by 25 microL of 0.6 M H2SO4, 1 h at 80 degrees C for acid hydrolysis of PS precedes the periodate treatment in the purpald assay. The concentration of the samples can be calculated from the sample absorbance and the reference standard curve constructed from the reference concentrations of the same PS (well-characterized) and their corresponding absorbance values assayed in the same plate. The purpald assay provides a tool in addition to the existing ones for the measurement of glycol-containing PS. Among the usefulness of this method are the determinations of the glycerol content in the phospho-glycerol-containing PS and the SG and UG contents and structural integrity in PS and conjugate vaccines.
repeating unit, bacterial polysaccharides, quantification
NCBI PubMed ID: 11520034Publication DOI: 10.1006/abio.2001.5230Journal NLM ID: 0370535Publisher: Academic Press
Institutions: Laboratory of Bacterial Polysaccharides, Division of Bacterial, Parasitic and Allergenic Products, OVRR, CBER, FDA, 8800 Rockville Pike, Bethesda, MD, USA
Methods: purpald assay measurement
- Article ID: 926
Lemercinier X, Jones C "Full 1H NMR assignment and detailed O-acetylation patterns of capsular polysaccharides from Neisseria meningitidis used in vaccine production" -
Carbohydrate Research 296(1-4) (1996) 83-96
We report esssentially complete 1H NMR assignments for the capsular polysaccharides from Neisseria meningitidis serotypes A, C, W-135, and Y. These polysaccharides are components of current polysaccharide vaccines against meningococcal infection and of the polysaccharide-protein conjugate vaccines under development. From these NMR data the pattern of O-acetylation was determined. O-Acetylation of the W-135 polysaccharide is reporded for the first time. We also show that, for the Types C and W-135 polysaccharides, a migration of O-acetyl groups occurs during storage in solution, and demonstrate that high field 1H NMR represents a simple and sensititve method to define the O-acetylation pattern of individual batches of these polysaccharides.
NMR, structure, Neisseria meningitidis, Neisseria, capsular, polysaccharide, capsular polysaccharide, capsular polysaccharides, polysaccharides, O-acetylation, production, vaccine, assignment, 1H NMR, pattern
Publication DOI: 10.1016/S0008-6215(96)00253-4Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: cjones@nibsc.ac.uk
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR-2D, NMR
- Article ID: 3570
Yongye AB, Gonzalez-Outeirino J, Glushka J, Schultheis V, Woods RJ "The Conformational Properties of Methyl α-(2,8)-Di/Trisialosides and Their N-Acyl Analogues: Implications for Anti-Neisseria meningitidis B Vaccine Design" -
Biochemistry 47(47) (2008) 12493-12514
The conformational properties of di- and trisaccharide fragments of the polysialic acid O-antigen capsular polysaccharide (CPS) of Neisseria meningitidis B (NmB) have been investigated by a combination of solution phase NMR spectroscopy and explicit-solvent molecular dynamics (MD) simulations. Simulations employing 100 ns of conventional MD, as well as 160 ns of replica exchange MD (REMD), with the GLYCAM06 force field were shown to be in agreement with experimental NMR scalar J-coupling and NOE values. The presence of conformational families has been determined by monitoring interglycosidic torsion angles, by comparing structural superimpositions, as well as via a Bayesian statistical analysis of the torsional data. Attempts to augment the immunogenicity of NmB CPS often involve chemical modifications of the N-acetyl moiety. Here the effects of these chemical group modifications on the conformational properties of the trisialoside have been probed via REMD simulations of the N-glycolyl, N-propionyl, N-propyl and N-butanoyl analogues. Although there were conformational families unique to each non-native analogue, the chemical modifications resulted in largely equivalent overall conformational phase-spaces compared to the native trisialoside. On the basis of the conformational distributions, these shared conformational properties suggest that a recurrent global conformational epitope may be present in both the native and chemically modified CPS fragments. Explanations are therefore provided for monoclonal antibody cross-reactivity, in terms of recognition of a shared global CPS conformation, as well as for lack of cross-reactivity, in terms of fine structural differences associated with the N-acyl groups, which may be dominant in highly matured antibody responses
NMR, conformation, Neisseria meningitidis, capsular polysaccharide, antibody response, vaccine
NCBI PubMed ID: 18954144Publication DOI: 10.1021/bi800431cJournal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: rwoods@ccrc.uga.edu
Institutions: Complex Carbohydrate Research Center, Athens, GA, USA, and Theoretische Biophysik, Lehrstuhl fur Biomolekulare Optik, Ludwig-Maximilians-Universitat, Oettingenstrasse 67, 80538 Munich, Germany
Methods: 13C NMR, 1H NMR, NMR-2D, 31P NMR, NMR-1D
- Article ID: 3664
Bergfeld AK, Claus H, Lorenzen NK, Spielmann F, Vogel U, Muhlenhoff M "The polysialic acid-specific O-acetyltransferase OatC from Neisseria meningitidis serogroup C evolved apart from other bacterial sialate O-acetyltransferases" -
Journal of Biological Chemistry 284(1) (2009) 6-16
Neisseria meningitidis serogroup C is a major cause of bacterial meningitis and septicaemia. This human pathogen is protected by a capsule composed of α2,9-linked polysialic acid that represents an important virulence factor. In the majority of strains, the capsular polysaccharide is modified by O-acetylation at C-7 or C-8 of the sialic acid residues. The gene encoding the capsule modifying O-acetyltransferase is part of the capsule gene complex and shares no sequence similarities with other proteins. Here, we describe the purification and biochemical characterization of recombinant OatC. The enzyme was found as a homodimer, with the first 34 amino acids forming an efficient oligomerization domain that worked even in a different protein context. Using acetyl-CoA as donor substrate, OatC transferred acetyl groups exclusively onto polysialic acid joined by α2,9-linkages and did not act on free or CMP-activated sialic acid. Motif scanning revealed a nucleophile elbow motif (GXS286XGG), which is a hallmark of α/β-hydrolase fold enzymes. In a comprehensive site-directed mutagenesis study, we identified a catalytic triad composed of Ser-286, Asp-376, and His-399. Consistent with a double-displacement mechanism common to α/β-hydrolase fold enzymes, a covalent acetylenzyme intermediate was found. Together with secondary structure prediction highlighting an α/β-hydrolase fold topology, our data provide strong evidence that OatC belongs to the α/β-hydrolase fold family. This clearly distinguishes OatC from all other bacterial sialate O-acetyltransferases known so far because these are members of the hexapeptide repeat family, a class of acyltransferases that adopt a left-handed beta-helix fold and assemble into catalytic trimers.
biosynthesis, Neisseria meningitidis, Neisseria, capsular polysaccharide, capsule, polysialic acid, acyltransferase, meningitis
NCBI PubMed ID: 18986988Publication DOI: 10.1074/jbc.M807518200Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: muehlenhoff.martina@mh-hannover.de
Institutions: Department of Cellular Chemistry, Medical School Hannover, 30623 Hannover, Germany
Methods: SDS-PAGE, genetic methods, biochemical methods, immunoblotting
- Article ID: 3727
Lee HJ, Rakic B, Gilbert M, Wakarchuk WW, Withers SG, Strynadka NC "Structural and kinetic characterizations of the polysialic acid O-acetyltransferase OatWY from Neisseria meningitidis" -
Journal of Biological Chemistry 284(36) (2009) 24501-24511
The neuroinvasive pathogen Neisseria meningitidis has 13 capsular serogroups, but the majority of disease is caused by only 5 of these. Groups B, C, Y, and W-135 all display a polymeric sialic acid-containing capsule that provides a means for the bacteria to evade the immune response during infection by mimicking host sialic acid-containing cell surface structures. These capsules in serogroups C, Y, and W-135 can be further acetylated by a sialic acid-specific O-acetyltransferase, a modification that correlates with decreased immunoreactivity and increased virulence. In N. meningitidis serogroup Y, the O-acetylation reaction is catalyzed by the enzyme OatWY, which we show has clear specificity toward the serogroup Y capsule ([Glc-(α1→4)-Sia](n)). To understand the underlying molecular basis of this process, we have performed crystallographic analysis of OatWY with bound substrate as well as determined kinetic parameters of the wild type enzyme and active site mutants. The structure of OatWY reveals an intimate homotrimer of left-handed beta-helix motifs that frame a deep active site cleft selective for the polysialic acid-bearing substrate. Within the active site, our structural, kinetic, and mutagenesis data support the role of two conserved residues in the catalytic mechanism (His-121 and Trp-145) and further highlight a significant movement of Tyr-171 that blocks the active site of the enzyme in its native form. Collectively, our results reveal the first structural features of a bacterial sialic acid O-acetyltransferase and provide significant new insight into its catalytic mechanism and specificity for the capsular polysaccharide of serogroup Y meningococci.
Neisseria meningitidis, virulence, O-acetylation, sialic acid, acetyltransferase, OatWY, serogroup Y
NCBI PubMed ID: 19525232Publication DOI: 10.1074/jbc.M109.006049Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: natalie@byron.biochem.ubc.ca
Institutions: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada
Methods: genetic methods, biochemical methods, molecular modeling, crystallization
- Article ID: 4430
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: 4509
Deng L, Chen X, Varki A "Exploration of sialic acid diversity and biology using sialoglycan microarrays" -
Biopolymers 99(10) (2013) 650-665
Sialic acids (Sias) are a group of alpha-keto acids with a nine-carbon backbone, which display many types of modifications in nature. The diversity of natural Sia presentations is magnified by a variety of glycosidic linkages to underlying glycans, the sequences and classes of such glycans, as well as the spatial organization of Sias with their surroundings. This diversity is closely linked to the numerous and varied biological functions of Sias. Relatively large libraries of natural and unnatural Sias have recently been chemically/chemoenzymatically synthesized and/or isolated from natural sources. The resulting sialoglycan microarrays have proved to be valuable tools for the exploration of diversity and biology of Sias. Here we provide an overview of Sia diversity in nature, the approaches used to generate sialoglycan microarrays, and the achievements and challenges arising.
diversity, sialic acids, sialoglycan microarrays
NCBI PubMed ID: 23765393Publication DOI: 10.1002/bip.22314Journal NLM ID: 0372525Publisher: Wiley Interscience
Correspondence: alvarki@ucsd.edu
Institutions: Departments of Medicine and Cellular & Molecular Medicine, Glycobiology Research and Training Center, University of California, San Diego, La Jolla, CA, 92093-0687, Department of Chemistry, University of California, Davis, CA 95616
- Article ID: 4608
Romanow A, Haselhorst T, Stummeyer K, Claus H, Bethe A, Muhlenhoff M, Vogel U, Von Itzstein M, Gerardy-Schahn R "Biochemical and biophysical characterization of the sialyl-/hexosyltransferase synthesizing the meningococcal serogroup w135 heteropolysaccharide capsule" -
Journal of Biological Chemistry 288(17) (2013) 11718-11730
Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis and sepsis. Crucial virulence determinants of pathogenic Nm strains are the polysaccharide capsules that support invasion by hindering complement attack. In NmW-135 and NmY the capsules are built from the repeating units (→6)-α-D-Gal-(1→4)-α-Neu5Ac-(2→)n and (→6)-α-D-Glc-(1→4)-α-Neu5Ac-(2→)n, respectively. These unusual heteropolymers represent unique examples of a conjugation between sialic acid and hexosyl-sugars in a polymer chain. Moreover, despite the various catalytic strategies needed for sialic acid and hexose transfer, single enzymes (SiaDW-135/Y) have been identified to form these heteropolymers. Here we used SiaDW-135 as a model system to delineate structure-function relationships. In size exclusion chromatography active SiaDW-135 migrated as a monomer. Fold recognition programs suggested two separate glycosyltransferase domains, both containing a GT-B-fold. Based on conserved motifs predicted folds could be classified as a hexosyl- and sialyltransferase. To analyze enzyme properties and interplay of the two identified glycosyltransferase domains, saturation transfer difference NMR and mutational studies were carried out. Simultaneous and independent binding of UDP-Gal and CMP-Sia was seen in the absence of an acceptor as well as when the catalytic cycle was allowed to proceed. Enzyme variants with only one functionality were generated by site-directed mutagenesis and shown to complement each other in trans when combined in an in vitro test system. Together the data strongly suggests that SiaDW-135 has evolved by fusion of two independent ancestral genes encoding sialyl- and galactosyltransferase activity.
biosynthesis, Neisseria meningitidis, sialic acid, glycosyltransferase, capsule polymerases, NmW-135
NCBI PubMed ID: 23439648Publication DOI: 10.1074/jbc.M113.452276Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: gerardy-schahn.rita@mh-hannover.de
Institutions: From the Institute for Cellular Chemistry, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany
Methods: SDS-PAGE, DNA techniques, genetic methods, radioactivity measurement, STD NMR
- Article ID: 4639
Willis LM, Whitfield C "Structure, biosynthesis, and function of bacterial capsular polysaccharides synthesized by ABC transporter-dependent pathways" -
Carbohydrate Research 378 (2013) 35-44
Bacterial capsules are formed primarily from long-chain polysaccharides with repeat-unit structures. A given bacterial species can produce a range of capsular polysaccharides (CPSs) with different structures and these help distinguish isolates by serotyping, as is the case with Escherichia coli K antigens. Capsules are important virulence factors for many pathogens and this review focuses on CPSs synthesized via ATP-binding cassette (ABC) transporter-dependent processes in Gram-negative bacteria. Bacteria utilizing this pathway are often associated with urinary tract infections, septicemia, and meningitis, and E. coli and Neisseria meningitidis provide well-studied examples. CPSs from ABC transporter-dependent pathways are synthesized at the cytoplasmic face of the inner membrane through the concerted action of glycosyltransferases before being exported across the inner membrane and translocated to the cell surface. A hallmark of these CPSs is a conserved reducing terminal glycolipid composed of phosphatidylglycerol and a poly-3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) linker. Recent discovery of the structure of this conserved lipid terminus provides new insights into the early steps in CPS biosynthesis.
biosynthesis, capsular polysaccharides, glycosyltransferases, gram negative bacteria, export, ABC transporters
NCBI PubMed ID: 23746650Publication DOI: 10.1016/j.carres.2013.05.007Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: C. Whitfield
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
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2. Compound ID: 2697
Structure type: polymer chemical repeating unit
Trivial name: type Y polysaccharide
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_158561,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 926
Lemercinier X, Jones C "Full 1H NMR assignment and detailed O-acetylation patterns of capsular polysaccharides from Neisseria meningitidis used in vaccine production" -
Carbohydrate Research 296(1-4) (1996) 83-96
We report esssentially complete 1H NMR assignments for the capsular polysaccharides from Neisseria meningitidis serotypes A, C, W-135, and Y. These polysaccharides are components of current polysaccharide vaccines against meningococcal infection and of the polysaccharide-protein conjugate vaccines under development. From these NMR data the pattern of O-acetylation was determined. O-Acetylation of the W-135 polysaccharide is reporded for the first time. We also show that, for the Types C and W-135 polysaccharides, a migration of O-acetyl groups occurs during storage in solution, and demonstrate that high field 1H NMR represents a simple and sensititve method to define the O-acetylation pattern of individual batches of these polysaccharides.
NMR, structure, Neisseria meningitidis, Neisseria, capsular, polysaccharide, capsular polysaccharide, capsular polysaccharides, polysaccharides, O-acetylation, production, vaccine, assignment, 1H NMR, pattern
Publication DOI: 10.1016/S0008-6215(96)00253-4Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: cjones@nibsc.ac.uk
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR-2D, NMR
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3. Compound ID: 2698
Structure type: polymer chemical repeating unit
Trivial name: type Y polysaccharide
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 926
Lemercinier X, Jones C "Full 1H NMR assignment and detailed O-acetylation patterns of capsular polysaccharides from Neisseria meningitidis used in vaccine production" -
Carbohydrate Research 296(1-4) (1996) 83-96
We report esssentially complete 1H NMR assignments for the capsular polysaccharides from Neisseria meningitidis serotypes A, C, W-135, and Y. These polysaccharides are components of current polysaccharide vaccines against meningococcal infection and of the polysaccharide-protein conjugate vaccines under development. From these NMR data the pattern of O-acetylation was determined. O-Acetylation of the W-135 polysaccharide is reporded for the first time. We also show that, for the Types C and W-135 polysaccharides, a migration of O-acetyl groups occurs during storage in solution, and demonstrate that high field 1H NMR represents a simple and sensititve method to define the O-acetylation pattern of individual batches of these polysaccharides.
NMR, structure, Neisseria meningitidis, Neisseria, capsular, polysaccharide, capsular polysaccharide, capsular polysaccharides, polysaccharides, O-acetylation, production, vaccine, assignment, 1H NMR, pattern
Publication DOI: 10.1016/S0008-6215(96)00253-4Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: cjones@nibsc.ac.uk
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR-2D, NMR
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4. Compound ID: 4161
Structure type: polymer chemical repeating unit
; >200000
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 1534
Lamb DH, Lei QP, Hakim N, Rizzo S, Cash P "Determination of meningococcal polysaccharides by capillary zone electrophoresis" -
Analytical Biochemistry 338(2) (2005) 263-269
Meningococcal polysaccharides are medically important molecules and are the active components of vaccines against Neisseria meningitidis serogroups A, C, W135, and Y. This study demonstrates that free solution capillary zone electrophoresis (CZE) using simple phosphate/borate separation buffers is capable of separating intact, native polysaccharides from these four serogroups. Separation appeared to be robust with respect to variations in test conditions and behaved in expected ways with respect to changes in temperature, ionic strength, and addition of an organic modifier. Serogroups W135 and Y are composed of sialic acid residues alternating with either galactose or glucose, respectively. Separation of these serogroups could be achieved using phosphate buffer and was therefore not dependent on differential complexation with borate. Addition of sodium dodecyl sulfate to the separation buffer (i.e., MEKC) resulted in peak splitting for all four serogroups. Changes in polysaccharide size did not affect migration time for the size range examined, but serogroup C polysaccharide (a sialic acid homopolymer) was separable from sialic acid monosaccharide. CZE quantification of multiple lots of each of the four serogroups was compared to wet chemical determination by phosphorus or sialic acid measurement. Results from CZE determination showed good agreement with the wet chemical methods
meningococcal, Neisseria, variation, polysaccharide, polysaccharides, group, molecule, determination, acid, phosphate, serogroup, chemical, method, sialic acid, vaccines, glucose, component, methods, galactose, time, measurement, monosaccharide, vaccine, solution, change, size, C-polysaccharide, quantification, separation, homopolymer, active, phosphorus, C polysaccharide, Capillary zone electrophoresis, organic, native, migration, electrophoresis, free, temperature, sulfate, development, test, multiple, borate, buffer, complexation, ionic, meningococcal polysaccharides, serogroup C, Sodium Dodecyl Sulfate, splitting
NCBI PubMed ID: 15745746Journal NLM ID: 0370535Publisher: Academic Press
Institutions: Analytical Development Group, Aventis Pasteur, Swiftwater, PA, USA
Methods: CE
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5. Compound ID: 11418
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 4606
Ravenscroft N, Wheeler JX, Jones C "Bioanalysis of meningococcal vaccines" -
Bioanalysis 2(2) (2010) 343-361
Meningococcal meningitis is feared because of the rapid onset of severe disease from mild symptoms and, therefore, is an important target for vaccine research. Five serogroups, defined by the structures of their capsular polysaccharides, are responsible for the vast majority of disease. Protection against four of these five serogroups can be obtained with polysaccharide or glycoconjugate vaccines, in which fragments of the capsular polysaccharides attached to a carrier protein generate anticarbohydrate immune responses, whilst protection against group B disease requires protein immunogens, often presented in vesicles containing outer membrane proteins. Glycoconjugate vaccines are now an established technology, but outer-membrane protein vaccines are still under development and present significant challenges. This review discusses physicochemical approaches to the characterization and quality control of these vaccines, as well as highlighting the problems and differences in vaccine design required for protection against different serogroups of the same species of pathogen.
conjugate vaccine, Meningococcal Vaccines, meningococcal meningitis
NCBI PubMed ID: 21083312Publication DOI: 10.4155/bio.09.181Journal NLM ID: 101512484Publisher: London: Future Science
Correspondence: Neil.Ravenscroft@uct.ac.za
Institutions: Bioanalytical and Vaccine Research, Department of Chemistry, University of Cape Town, Rondebosch 7701, South Africa, Laboratory for Molecular Structure, National Institute for Biological Standards and Control, Health Protection Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK
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6. Compound ID: 12151
|
/Variants 0/-+
|
-6)-a-D-Glcp-(1-4)-a-Neup5Ac-(2-
/Variants 0/ is:
?%Ac-9)-
OR (exclusively)
?%Ac-7)- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_158561,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 4835
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: 5015
Kuttel MM, Timol Z, Ravenscroft N "Cross-protection in Neisseria meningitidis serogroups Y and W polysaccharides: A comparative conformational analysis" -
Carbohydrate Research (2017) 40-47
The capsular polysaccharide is the main virulence factor in meningococcus. The capsular polysaccharides for meningococcal serogroups Y and W are almost identical polymers of hexose-sialic acid, suggesting the possibility of cross-protection between group Y and W vaccines. However, early studies indicated that they elicit different levels of cross-protection. Here we explore the conformations of the meningococcal Y and W polysaccharides with molecular dynamics simulations of three repeating unit oligosaccharide strands. We find differences in Y and W antigen conformation: the Y polysaccharide has a single dominant conformation, whereas W exhibits a family of conformations including the Y conformation. This result is supported by our NMR NOESY analysis, which indicates key close contacts for W that are not present in Y. These conformational differences provide an explanation for the different levels of cross-protection measured for the Y and W monovalent vaccines and the high group W responses observed in HibMenCY-TT vaccinees.
molecular modelling, Meningococcus, Neisseria meningitidis, capsular polysaccharide, carbohydrate antigens, cross-protection, molecular dynamics simulation
NCBI PubMed ID: 28501716Publication DOI: 10.1016/j.carres.2017.05.004Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mkuttel@cs.uct.ac.za
Institutions: Department of Computer Science, University of Cape Town, Cape Town, 7701, South Africa
Methods: 13C NMR, 1H NMR, NMR-2D, conformation analysis, MD simulations, de-O-acetylation
- Article ID: 5128
Berti F, De Ricco R, Rappuoli R "Role of O-Acetylation in the Immunogenicity of Bacterial Polysaccharide Vaccines" -
Molecules 23(6) (2018) pii E1340
The incidence of infectious diseases caused by several bacterial pathogens such as Haemophilus influenzae type b, Streptococcus pneumoniae, and Neisseria meningitidis, has been dramatically reduced over the last 25 years through the use of glycoconjugate vaccines. The structures of the bacterial capsular polysaccharide (CPS) antigens, extracted and purified from microbial cultures and obtained with very high purity, show that many of them are decorated by O-acetyl groups. While these groups are often considered important for the structural identity of the polysaccharides, they play a major role in the functional immune response to some vaccines such as meningococcal serogroup A and Salmonella typhi Vi, but do not seem to be important for many others, such as meningococcal serogroups C, W, Y, and type III Group B Streptococcus. This review discusses the O-acetylation status of CPSs and its role in the immunological responses of these antigens.
O-acetylation, Bacterial polysaccharide, conjugate vaccines, Bacterial Vaccines, carbohydrate antigens
NCBI PubMed ID: 29865239Publication DOI: 10.3390/molecules23061340Journal NLM ID: 100964009Publisher: Basel, Switzerland: MDPI
Correspondence: rino.x.rappuoli@gsk.com
Institutions: External R&D, GSK Vaccines, 53100 Siena, Italy
- Article ID: 5161
Hlozek J, Kuttel MM, Ravenscroft N "Conformations of Neisseria meningitidis serogroup A and X polysaccharides: The effects of chain length and O-acetylation" -
Carbohydrate Research 465 (2018) 44-51
Neisseria meningitidis is a major cause of bacterial meningitis worldwide especially in Africa. The capsular polysaccharide (CPS) is the main virulence factor and the target antigen for polysaccharide and conjugate vaccines. The high burden of serogroup A disease in the Meningitis Belt of sub-Saharan Africa led to the introduction of MenAfriVac®, which has successfully reduced the number of cases of group A disease. However, several outbreaks caused by other serogroups have been reported, including those due to serogroup X. The capsular polysaccharides of serogroups A and X are both homopolymers of amino sugars (?-D-ManNAc and ?-D-GlcNAc) containing phosphodiester linkages at C-6 and C-4, respectively. The similarity of the primary structures of the two polysaccharides suggests that serogroup A vaccination may provide cross-protection against serogroup X disease. Molecular dynamics simulations of a series of serogroup A and X oligosaccharides reveal that the MenA CPS behaves as a flexible random coil which becomes less conformationally defined as the length increases, whereas serogroup X forms a more stable regular helical structure. The presence of the MenX helix is supported by NMR analysis; it has four residues per turn and becomes more stable as the chain length increases. Licensed MenA vaccines are largely O-acetylated at C-3: simulations show that these O-acetyl groups are highly solvent exposed and their presence favors more extended conformations compared to the more compact conformations of MenA without O-acetylation. These findings may have implications for the design of optimal conjugate vaccines.
Neisseria meningitidis, capsular polysaccharide, molecular modeling, Antigen conformation
NCBI PubMed ID: 29940397Publication DOI: 10.1016/j.carres.2018.06.007Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: neil.ravenscroft@uct.ac.za
Institutions: Department of Chemistry, University of Cape Town, Rondebosch, 7701, South Africa, Department of Computer Science, University of Cape Town, Rondebosch, 7701, South Africa
Methods: conformation analysis, MD simulations, post-simulation analysis
- Article ID: 5774
Gomez-Redondo M, Arda A, Gimeno A, Jiménez-Barbero J "Bacterial polysaccharides: conformation, dynamics and molecular recognition by antibodies" -
Drug Discovery Today: Technologies (2020) 1-11
Bacterial infections are the cause of different severe health conditions and new therapies to combat these pathogens have been widely investigated. Carbohydrates, being complex structures covering the surface of bacteria, are considered relevant targets for antibody and vaccine development. The biological activities in pathogenesis of bacterial capsular polysaccharides and lipopolisaccharides and their unique structures have boosted the study of the minimal antigenic binding epitopes and the structural details of antibody-carbohydrate recognition. This review describes the most recent advances on the field, examining the structure, conformation and dynamics of relevant bacterial carbohydrates and their complexes with antibodies. The understanding of key factors governing the recognition process is fundamental for the progress toward the development of specific and efficient bacterial therapeutics.
conformation, structure, capsular polysaccharides, epitope, recognition, dynamics, binding, vaccine, bacterial infection
NCBI PubMed ID: 33388123Publication DOI: 10.1016/j.ddtec.2020.08.002Journal NLM ID: 101235076Publisher: Oxford: Elsevier
Institutions: CIC bioGUNE, Basque Research Technology Alliance, BRTA, Bizkaia Technology Park, Building 800, 48160 Derio, Spain, Ikerbasque, Basque Foundation for Science, 48009 Bilbao, Spain, Department Organic Chemistry II, Faculty of Science and technology, UPV-EHU, 48940 Leioa, Spain
- Article ID: 6032
Berti F "NMR characterization of a multi-valent conjugate vaccine against Neisseria meningitidis A, C, W, Y and Haemophilus influenzae b infections" -
Journal of Pharmaceutical and Biomedical Analysis 205 (2021) 114302
Physicochemical technologies are a powerful tool for the structural characterization of vaccine antigens both at bulk level as well as on the final formulation. High-field Nuclear Magnetic Resonance (NMR) spectroscopy has been found to be an extremely and robust tool for tracking the industrial process manufacturing of carbohydrate-based vaccines. I have applied NMR spectroscopy to the characterization of a penta-valent conjugate vaccine against Neisseria meninigitidis group A, C, W, Y (MenACWY) and Haemophilus influenzae type b (Hib) infections, constituted of capsule derived polysaccharide fragments independently conjugated to CRM197 protein carrier (CRM-MenA, CRM-MenC, CRM-MenW, CRM-MenY, CRM-Hib). 1H NMR has been used for the identity testing of the carbohydrate antigens and of the vaccine formulation. The application of NMR-based assays on multivalent conjugate vaccines looks to be a promising approach for identity and stability analyses useful for future vaccines development.
glycoconjugates, nuclear magnetic resonance spectroscopy, vaccines, Haemophilus influenzae type b, Bacterial Infections, Neisseria meningitidis serogroup A/C/W/Y
NCBI PubMed ID: 34388671Publication DOI: 10.1016/j.jpba.2021.114302Journal NLM ID: 8309336Publisher: London: Elsevier
Correspondence: francesco.x.berti@gsk.com
Institutions: GSK, Via Fiorentina 1, 53100 Siena, Italy
Methods: 13C NMR, 1H NMR, 31P NMR, vaccine formulations
- Article ID: 6096
Kuttel MM, Berti F, Ravenscroft N "Molecular modeling provides insights into the loading of sialic acid-containing antigens onto CRM 197: the role of chain flexibility in conjugation efficiency and glycoconjugate architecture" -
Glycoconjugate Journal 38(4) (2021) 411-419
Vaccination is the most cost-effective way to control disease caused by encapsulated bacteria; the capsular polysaccharide (CPS) is the primary virulence factor and vaccine target. Neisseria meningitidis (Nm) serogroups B, C, Y and W all contain sialic acid, a common surface feature of human pathogens. Two protein-based vaccines against serogroup B infection are available for human use while four tetravalent conjugate vaccines including serogroups C, W and Y have been licensed. The tetravalent Menveo® conjugate vaccine is well-defined: a simple monomeric structure of oligosaccharides terminally conjugated to amino groups of the carrier protein CRM197. However, not only is there a surprisingly low limit for antigen chain attachment to CRM197, but different serogroup saccharides have consistently different CRM197 loading, the reasons for which are unclear. Understanding this phenomenon is important for the long-term goal of controlling conjugation to prepare conjugate vaccines of optimal immunogenicity. Here we use molecular modeling to explore whether antigen flexibility can explain the varying antigen loading of the conjugates. Because flexibility is difficult to separate from other structural factors, we focus on sialic-acid containing CPS present in current glycoconjugate vaccines: serogroups NmC, NmW and NmY. Our simulations reveal a correlation between Nm antigen flexibility (NmW > NmC > NmY) and the number of chains attached to CRM197, suggesting that increased flexibility enables accommodation of additional chains on the protein surface. Further, in silico models of the glycoconjugates confirm the relatively large hydrodynamic size of the saccharide chains and indicate steric constraints to further conjugation.
Neisseria meningitidis, capsular polysaccharide, molecular modeling, Antigen conformation, conjugation efficiency, glycoconjugate structure
NCBI PubMed ID: 33721150Publication DOI: 10.1007/s10719-021-09991-xJournal NLM ID: 8603310Publisher: Kluwer Academic Publishers
Correspondence: neil.ravenscroft@uct.ac.za
Institutions: Department of Computer Science, University of Cape Town, 7701, Cape Town, South Africa, Technical Research & Development, GSK Vaccines S.r.l, Via Fiorentina 1, 53100, Siena, Italy, Department of Chemistry, University of Cape Town, 7701, Cape Town, South Africa
Methods: MD simulations, molecular modeling, CarbBuilder, CHARMM36
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7. Compound ID: 12227
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_158561,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 4867
Tzeng YL, Thomas J, Stephens DS "Regulation of capsule in Neisseria meningitidis" -
Critical Reviews in Microbiology 42(5) (2016) 759-772
Neisseria meningitidis, a devastating pathogen exclusive to humans, expresses capsular polysaccharides that are the major meningococcal virulence determinants and the basis for successful meningococcal vaccines. With rare exceptions, the expression of capsule (serogroups A, B, C, W, X, Y) is required for systemic invasive meningococcal disease. Changes in capsule expression or structure (e.g. hypo- or hyper-encapsulation, capsule 'switching', acetylation) can influence immunologic diagnostic assays or lead to immune escape. The loss or down-regulation of capsule is also critical in meningococcal biology facilitating meningococcal attachment, microcolony formation and the carriage state at human mucosal surfaces. Encapsulated meningococci contain a cps locus with promoters located in an intergenic region between the biosynthesis and the conserved capsule transport operons. The cps intergenic region is transcriptionally regulated (and thus the amount of capsule expressed) by IS element insertion, by a two-component system, MisR/MisS and through sequence changes that result in post-transcriptional RNA thermoregulation. Reversible on-off phase variation of capsule expression is controlled by slipped strand mispairing of homo-polymeric tracts and by precise insertion and excision of IS elements (e.g. IS1301) in the biosynthesis operon. Capsule structure can be altered by phase-variable expression of capsular polymer modification enzymes or 'switched' through transformation and homologous recombination of different polymerases. Understanding the complex regulation of meningococcal capsule has important implications for meningococcal biology, pathogenesis, diagnostics, current and future vaccine development and vaccine strategies.
Neisseria meningitidis, capsular polysaccharide, Bacterial virulence regulation, capsule switching, meningococcal diseases
NCBI PubMed ID: 26089023Publication DOI: 10.3109/1040841X.2015.1022507Journal NLM ID: 8914274Publisher: CRC Press
Correspondence: dstep01@emory.edu
Institutions: Department of Medicine, Emory University School of Medicine, Woodruff Health Sciences Center, Atlanta, GA, USA
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8. Compound ID: 16287
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_136794,IEDB_142488,IEDB_144998,IEDB_146100,IEDB_146664,IEDB_148489,IEDB_149174,IEDB_158547,IEDB_158561,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_84
The structure is contained in the following publication(s):
- Article ID: 6318
Srtefanetti G, Maclennan CA, Micoli F "Impact and Control of Sugar Size in Glycoconjugate Vaccines" -
Molecules 27(19) (2022) 6432
Glycoconjugate vaccines have contributed enormously to reducing and controlling encapsulated bacterial infections for over thirty years. Glycoconjugate vaccines are based on a carbohydrate antigen that is covalently linked to a carrier protein; this is necessary to cause T cell responses for optimal immunogenicity, and to protect young children. Many interdependent parameters affect the immunogenicity of glycoconjugate vaccines, including the size of the saccharide antigen. Here, we examine and discuss the impact of glycan chain length on the efficacy of glycoconjugate vaccines and report the methods employed to size polysaccharide antigens, while highlighting the underlying reaction mechanisms. A better understanding of the impact of key parameters on the immunogenicity of glycoconjugates is critical to developing a new generation of highly effective vaccines.
glycoconjugates, vaccine, immunogenicity, fragmentation of polysaccharides, sugar length
NCBI PubMed ID: 36234967Publication DOI: 10.3390/molecules27196432Journal NLM ID: 100964009Publisher: Basel, Switzerland: MDPI
Correspondence: G. Srtefanetti
Institutions: Department of Biomolecular Sciences, University of Urbino Carlo Bo, 61029 Urbino, Italy, Enteric and Diarrheal Diseases, Global Health, Bill & Melinda Gates Foundation, 500 5th Ave. N, Seattle, WA 98109, USA, The Jenner Institute, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, UK, The Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham B15 2TT, UK, GSK Vaccines Institute for Global Health, 53100 Siena, Italy
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