Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130646,IEDB_130650,IEDB_130670,IEDB_130697,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_1391962,IEDB_140088,IEDB_140108,IEDB_140122,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142488,IEDB_143794,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_232584,IEDB_885811,IEDB_983931,SB_137,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_29,SB_30,SB_7,SB_88

• Article ID: 1108
  Phillips NJ, Miller TJ, Engstrom JJ, Melaugh W, McLaughlin R, Apicella MA, Gibson BW "Characterization of chimeric lipopolysaccharides from Escherichia coli strain JM109 transformed with lipooligosaccharide synthesis genes (lsg) from Haemophilus influenzae" - Journal of Biological Chemistry 275(7) (2000) 4747-4758
  

  Previously, we reported the expression of chimeric lipopolysaccharides (LPS) in Escherichia coli strain JM109 (a K-12 strain) transformed with plasmids containing Haemophilus influenzae lipooligosaccharide synthesis genes (lsg) (Abu Kwaik, Y., McLaughlin, R. E., Apicella, M. A., and Spinola, S. M. (1991) Mol. Microbiol. 5, 2475-2480). In this current study, we have analyzed the O-deacylated LPS and free oligosaccharides from three transformants (designated pGEMLOS-4, pGEMLOS-5, and pGEMLOS-7) by matrix-assisted laser desorption ionization, electrospray ionization, and tandem mass spectrometry techniques, along with composition and linkage analyses. These data show that the chimeric LPS consist of the complete E. coli LPS core structure glycosylated on the 7-position of the non-reducing terminal branch heptose with oligosaccharides from H. influenzae. In pGEMLOS-7, the disaccharide Gal 1→3 GlcNAc1→ is added, and in pGEMLOS-5, the structure is extended to Gal 1→4 GlcNAc 1→3 Gal 1→3 GlcNAc1→. PGEMLOS-5 LPS reacts positively with monoclonal antibody 3F11, an antibody that recognizes the terminal disaccharide of lacto-N-neotetraose. In pGEMLOS-4 LPS, the 3F11 epitope is apparently blocked by glycosylation on the 6-position of the terminal Gal with either Gal or GlcNAc. The biosynthesis of these chimeric LPS was found to be dependent on a functional wecA (formerly rfe) gene in E. coli. By using this carbohydrate expression system, we have been able to examine the functions of the lsg genes independent of the effects of other endogenous Haemophilus genes and expressed proteins.

  Lipopolysaccharide, synthesis, Haemophilus, Haemophilus influenzae, lipopolysaccharides, Lipooligosaccharide, gene, strain, characterization, Escherichia, Escherichia coli

  NCBI PubMed ID: 10671507
  Publication DOI: 10.1074/jbc.275.7.4747
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: gibson@socrates.cgl.ucsf.edu
  Institutions: Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, USA, Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, Department of Microbiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
  Methods: ESI-MS, MALDI-MS, MS/MS, composition analysis, linkage analysis
  
   
  
  Escherichia coli pGEMLOS-4
  CSDB ID 4175 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130646,IEDB_130650,IEDB_130670,IEDB_130697,IEDB_134624,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_1391962,IEDB_140088,IEDB_140108,IEDB_140122,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142488,IEDB_143794,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_153201,IEDB_156493,IEDB_167070,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_232584,IEDB_742248,IEDB_983931,SB_137,SB_163,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_29,SB_30,SB_7,SB_88

• Article ID: 1108
  Phillips NJ, Miller TJ, Engstrom JJ, Melaugh W, McLaughlin R, Apicella MA, Gibson BW "Characterization of chimeric lipopolysaccharides from Escherichia coli strain JM109 transformed with lipooligosaccharide synthesis genes (lsg) from Haemophilus influenzae" - Journal of Biological Chemistry 275(7) (2000) 4747-4758
  

  Previously, we reported the expression of chimeric lipopolysaccharides (LPS) in Escherichia coli strain JM109 (a K-12 strain) transformed with plasmids containing Haemophilus influenzae lipooligosaccharide synthesis genes (lsg) (Abu Kwaik, Y., McLaughlin, R. E., Apicella, M. A., and Spinola, S. M. (1991) Mol. Microbiol. 5, 2475-2480). In this current study, we have analyzed the O-deacylated LPS and free oligosaccharides from three transformants (designated pGEMLOS-4, pGEMLOS-5, and pGEMLOS-7) by matrix-assisted laser desorption ionization, electrospray ionization, and tandem mass spectrometry techniques, along with composition and linkage analyses. These data show that the chimeric LPS consist of the complete E. coli LPS core structure glycosylated on the 7-position of the non-reducing terminal branch heptose with oligosaccharides from H. influenzae. In pGEMLOS-7, the disaccharide Gal 1→3 GlcNAc1→ is added, and in pGEMLOS-5, the structure is extended to Gal 1→4 GlcNAc 1→3 Gal 1→3 GlcNAc1→. PGEMLOS-5 LPS reacts positively with monoclonal antibody 3F11, an antibody that recognizes the terminal disaccharide of lacto-N-neotetraose. In pGEMLOS-4 LPS, the 3F11 epitope is apparently blocked by glycosylation on the 6-position of the terminal Gal with either Gal or GlcNAc. The biosynthesis of these chimeric LPS was found to be dependent on a functional wecA (formerly rfe) gene in E. coli. By using this carbohydrate expression system, we have been able to examine the functions of the lsg genes independent of the effects of other endogenous Haemophilus genes and expressed proteins.

  Lipopolysaccharide, synthesis, Haemophilus, Haemophilus influenzae, lipopolysaccharides, Lipooligosaccharide, gene, strain, characterization, Escherichia, Escherichia coli

  NCBI PubMed ID: 10671507
  Publication DOI: 10.1074/jbc.275.7.4747
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: gibson@socrates.cgl.ucsf.edu
  Institutions: Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, USA, Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, Department of Microbiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104
  Methods: ESI-MS, MALDI-MS, MS/MS, composition analysis, linkage analysis
  
   
  
  Escherichia coli pGEMLOS-4
  CSDB ID 4176 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130646,IEDB_130697,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_1391966,IEDB_140087,IEDB_140088,IEDB_140089,IEDB_140090,IEDB_140108,IEDB_140110,IEDB_140122,IEDB_141794,IEDB_141807,IEDB_142351,IEDB_142487,IEDB_142488,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_149144,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,IEDB_2189047,IEDB_419428,IEDB_419429,IEDB_983931,SB_145,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_30,SB_6,SB_7,SB_88

• Article ID: 1132
  Ram S, Cox AD, Wright JC, Vogel U, Getzlaff S, Boden R, Li J, Plested JS, Meri S, Gulati S, Stein DC, Richards JC, Moxon ER, Rice PA "Neisserial lipooligosaccharide is a target for complement component C4b: Inner core phosphoethanolamine residues define C4b linkage specificity" - Journal of Biological Chemistry 278(51) (2003) 50853-50862
  

  We identified Neisseria meningitidis lipooligosaccharide (LOS) as an acceptor for complement component C4b (C4b). Phosphoethanolamine (PEA) residues on the second heptose (HepII) residue in the LOS core structure formed amide linkages with C4b. PEA on the 6-position of HepII (6-PEA) was more efficient than 3-PEA in binding C4b. Strains bearing 6-PEA bound more C4b than strains with 3-PEA and were more susceptible to complement-mediated killing in serum bactericidal assays. Deleting 3-PEA from a strain that expressed both 3- and 6-PEA simultaneously on HepII did not decrease C4b binding. Glycose chain extension of the first heptose residue (HepI) influenced the nature of the C4b-LOS linkage. Predominantly ester C4b-LOS bonds were seen when lacto-N-neotetraose formed the terminus of the glycose chain extension of HepI with 3-PEA on HepII in the LOS core. Related LOS species with more truncated chain extensions from HepI bound C4b via amide linkages to 3-PEA on HepII. However, 6-PEA in the LOS core bound C4b even when the glycose chain from HepI bore lacto-N-neotetraose at the terminus. The C4A isoform exclusively formed amide linkages, while C4B bound meningococci preferentially via ester linkages. These data may serve to explain the preponderance of 3-PEA bearing meningococci among clinical isolates because 6-PEA enhances C4b binding that may facilitate clearance of 6-PEA-bearing strains resulting from enhanced serum killing by the classical pathway of complement

  structure, core, heptose, Lipooligosaccharide, Neisseria meningitidis, clinical, disease, isolate, LOS, meningococci, Neisseria, strain, chain, enhanced, infectious disease, specificity, linkage, inner core, medicine, phosphoethanolamine, Infectious, component, binding, lacto-N-neotetraose, acceptor, pathway, serum, bound, bactericidal, serum killing, species, killing, assay, decrease, terminus, amide, target, classical, clearance, complement, ester, pea

  NCBI PubMed ID: 14525973
  Publication DOI: 10.1074/jbc.M308364200
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: sram@bu.edu
  Institutions: Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A 0R6, Canada, Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742, Section of Infectious Diseases, Evans Biomedical Research Center, Boston University Medical Center, Boston, Massachusetts 02118, Molecular Infectious Diseases Group, Oxford University Department of Pediatrics, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom, nstitute for Hygiene and Microbiology, University of Wuerzburg, 97080 Wuerzburg, Germany, Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland-00014
  Methods: 1H NMR, ESI-MS, Western blotting
  
   
  
  Neisseria meningitidis MC51, Neisseria meningitidis 2120
  CSDB ID 4422 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130646,IEDB_130697,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_1391966,IEDB_140087,IEDB_140088,IEDB_140089,IEDB_140090,IEDB_140108,IEDB_140110,IEDB_140122,IEDB_141794,IEDB_141807,IEDB_142351,IEDB_142487,IEDB_142488,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_149144,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,IEDB_2189047,IEDB_419429,IEDB_419431,IEDB_983931,SB_145,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_30,SB_6,SB_7,SB_88

• Article ID: 1132
  Ram S, Cox AD, Wright JC, Vogel U, Getzlaff S, Boden R, Li J, Plested JS, Meri S, Gulati S, Stein DC, Richards JC, Moxon ER, Rice PA "Neisserial lipooligosaccharide is a target for complement component C4b: Inner core phosphoethanolamine residues define C4b linkage specificity" - Journal of Biological Chemistry 278(51) (2003) 50853-50862
  

  We identified Neisseria meningitidis lipooligosaccharide (LOS) as an acceptor for complement component C4b (C4b). Phosphoethanolamine (PEA) residues on the second heptose (HepII) residue in the LOS core structure formed amide linkages with C4b. PEA on the 6-position of HepII (6-PEA) was more efficient than 3-PEA in binding C4b. Strains bearing 6-PEA bound more C4b than strains with 3-PEA and were more susceptible to complement-mediated killing in serum bactericidal assays. Deleting 3-PEA from a strain that expressed both 3- and 6-PEA simultaneously on HepII did not decrease C4b binding. Glycose chain extension of the first heptose residue (HepI) influenced the nature of the C4b-LOS linkage. Predominantly ester C4b-LOS bonds were seen when lacto-N-neotetraose formed the terminus of the glycose chain extension of HepI with 3-PEA on HepII in the LOS core. Related LOS species with more truncated chain extensions from HepI bound C4b via amide linkages to 3-PEA on HepII. However, 6-PEA in the LOS core bound C4b even when the glycose chain from HepI bore lacto-N-neotetraose at the terminus. The C4A isoform exclusively formed amide linkages, while C4B bound meningococci preferentially via ester linkages. These data may serve to explain the preponderance of 3-PEA bearing meningococci among clinical isolates because 6-PEA enhances C4b binding that may facilitate clearance of 6-PEA-bearing strains resulting from enhanced serum killing by the classical pathway of complement

  structure, core, heptose, Lipooligosaccharide, Neisseria meningitidis, clinical, disease, isolate, LOS, meningococci, Neisseria, strain, chain, enhanced, infectious disease, specificity, linkage, inner core, medicine, phosphoethanolamine, Infectious, component, binding, lacto-N-neotetraose, acceptor, pathway, serum, bound, bactericidal, serum killing, species, killing, assay, decrease, terminus, amide, target, classical, clearance, complement, ester, pea

  NCBI PubMed ID: 14525973
  Publication DOI: 10.1074/jbc.M308364200
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: sram@bu.edu
  Institutions: Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A 0R6, Canada, Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742, Section of Infectious Diseases, Evans Biomedical Research Center, Boston University Medical Center, Boston, Massachusetts 02118, Molecular Infectious Diseases Group, Oxford University Department of Pediatrics, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom, nstitute for Hygiene and Microbiology, University of Wuerzburg, 97080 Wuerzburg, Germany, Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland-00014
  Methods: 1H NMR, ESI-MS, Western blotting
  
   
  
  Neisseria meningitidis 89I
  CSDB ID 4424 (all data & tools)

Show legend Show as text

Structure type: structural motif or average structure
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_114703,IEDB_120354,IEDB_123890,IEDB_130646,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_1391962,IEDB_140108,IEDB_140122,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142488,IEDB_143794,IEDB_144998,IEDB_144999,IEDB_145003,IEDB_146664,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,IEDB_241103,IEDB_241107,IEDB_241118,IEDB_885811,IEDB_983931,SB_137,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_29,SB_30,SB_7,SB_88

• Article ID: 2969
  Gmeiner J "The ribitol-phosphate-containing lipopolysaccharide from Proteus mirabilis, strain D52. Investigations of O-specific chains" - European Journal of Biochemistry 74 (1977) 171-180
  

  A soluble hydrophilic lipopolysaccharide, termed lipopolysaccharide II, isolated from Proteus mirabilis, strain D52 contained N-acetylglucosamine, glucose, galactose, ribitol phosphate and ethanolamine phosphate as constituents of the O-specific polysaccharide. Periodate oxidation studies were carried out on the polymer before and after dephosphorylation with hydrofluoric acid and on oligosaccharides derived from the polymer by partial acid hydrolysis. The results obtained indicate that the polysaccharide chain consists of the chemical repeating unit Gal-1,3(4)-GlcNAc-1,3-Glc-1,3-GlcNAc-, where GlcNAc stands for N-acetylglucosamine. Whereas the galactose residue is substituted at C-3 by ribitol phosphate, the glucose is substituted by ethanolamine phosphate at C-6.

  NCBI PubMed ID: 323005
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  
   
  
  Proteus mirabilis D52
  CSDB ID 122439 (all data & tools)

Show legend Show as text

Structure type: polymer chemical repeating unit
Compound class: EPS
Contained glycoepitopes: IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_142487,IEDB_142488,IEDB_145003,IEDB_146664,IEDB_149139,IEDB_151528,IEDB_151531,IEDB_153543,IEDB_167070,IEDB_190606,IEDB_983931,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_6,SB_7,SB_88

• Article ID: 3405
  Gorska S, Grycko P, Rybka J, Gamian A "Exopolysaccharides of lactic acid bacteria: structure and biosynthesis" - Postȩpy Higieny i Medycyny Doświadczalnej [Polish] 61 (2007) 805-818
  

  The group of lactic acid bacteria (LABs) includes four genera: Lactobacillus, Leuconostoc, Pediococcus, and Streptococcus. The most characteristic feature of this group of microorganisms is the production of lactic acid as a main product of carbohydrate metabolism. LABs are responsible for the fermentation of alimentary products and they also produce a variety of agents, among them exopolysaccharides (EPSs), which inhibit the growth of pathogenic bacteria. In this article on the different types of EPSs produced by LABs, data concerning their structure and biosynthesis are presented

  biosynthesis, structure, Streptococcus, Lactic acid bacteria, exopolysaccharides, Lactococcus, Lactobacillus, Leuconostoc

  NCBI PubMed ID: 18097339
  Journal NLM ID: 0421052
  Publisher: Warszawa: Panstwowy Zaklad Wydawnictw Lekarskich
  Institutions: Laboratorium Mikrobiologii Lekarskiej, Instytut Immunologii i Terapii Doswiadczalnej PAN im. L. Hirszfelda we Wroclawiu
  
   
  
  Lactobacillus helveticus TY 1-2
  CSDB ID 21992 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130646,IEDB_130650,IEDB_130654,IEDB_130655,IEDB_130697,IEDB_135813,IEDB_136044,IEDB_136045,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_140088,IEDB_140108,IEDB_140122,IEDB_141500,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144556,IEDB_144562,IEDB_144998,IEDB_145003,IEDB_145669,IEDB_146664,IEDB_147455,IEDB_149557,IEDB_150092,IEDB_150939,IEDB_151528,IEDB_151531,IEDB_152214,IEDB_158550,IEDB_167070,IEDB_174333,IEDB_190606,IEDB_2151203,IEDB_2189046,IEDB_2189047,IEDB_461720,IEDB_952752,IEDB_983931,SB_157,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_30,SB_7,SB_86,SB_88

• Article ID: 3520
  Moran AP "Relevance of fucosylation and Lewis antigen expression in the bacterial gastroduodenal pathogen Helicobacter pylori" - Carbohydrate Research 343(12) (2008) 1952-1965
  

  Helicobacter pylori is a prevalent bacterial, gastroduodenal pathogen of humans that can express Lewis (Le) and related antigens in the O-chains of its surface lipopolysaccharide. The O-chains of H. pylori are commonly composed of internal Le(x) units with terminal Le(x) or Le(y) units or, in some strains, with additional units of Le(a), Le(b), Le(c), sialyl-Le(x) and H-1 antigens, as well as blood groups A and B, thereby producing a mosaicism of antigenic units expressed. The genetic determination of the Le antigen biosynthetic pathways in H. pylori has been studied, and despite striking functional similarity, low sequence homology occurs between the bacterial and mammalian α(1,3/4)- and α(1,2)-fucosyltransferases. Factors affecting Le antigen expression in H. pylori, that can influence the biological impact of this molecular mimicry, include regulation of fucosyltransferase genes through slipped-strand mispairing, the activity and expression levels of the functional enzymes, the preferences of the expressed enzyme for distinctive acceptor molecules and the availability of activated sugar intermediates. Le mimicry was initially implicated in immune evasion and gastric adaptation by the bacterium, but more recent studies show a role in gastric colonization and bacterial adhesion with galectin-3 identified as the gastric receptor for polymeric Le(x) on the bacterium. From the host defence aspect, innate immune recognition of H. pylori by surfactant protein D is influenced by the extent of LPS fucosylation. Furthermore, Le antigen expression affects both the inflammatory response and T-cell polarization that develops after infection. Although controversial, evidence suggests that long-term H. pylori infection can induce autoreactive anti-Le antibodies cross-reacting with the gastric mucosa, in part leading to the development of gastric atrophy. Thus, Le antigen expression and fucosylation in H. pylori have multiple biological effects on pathogenesis and disease outcome.

  molecular mimicry, Helicobacter pylori, Fucosyltransferases, bacterial pathogenesis, Lewis antigens, fucosylation

  NCBI PubMed ID: 18279843
  Publication DOI: 10.1016/j.carres.2007.12.012
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: anthony.moran@nuigalway.ie
  Institutions: Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland, Institute for Glycomics, Gold Coast Campus, Griffith University, Queensland 4222, Australia
  Methods: NMR, sugar analysis, MS, genetic methods
  
   
  
  Helicobacter pylori NCTC 11637
  CSDB ID 23178 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130646,IEDB_130650,IEDB_130670,IEDB_130697,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137776,IEDB_1391962,IEDB_140088,IEDB_140108,IEDB_140122,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142488,IEDB_143794,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_232584,IEDB_983931,SB_137,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_29,SB_30,SB_7,SB_88

• Article ID: 3866
  Johansen EB, Szoka FC, Zaleski A, Apicella MA, Gibson BW "Utilizing the O-antigen Lipopolysaccharide Biosynthesis Pathway in Escherichia coli to Interrogate the Substrate Specificities of Exogenous Glycosyltransferase Genes in a Combinatorial Approach" - Glycobiology 20(6) (2010) 763-774
  

  In previous work, our laboratory generated novel chimeric lipopolysaccharides (LPS) in Escherichia coli transformed with a plasmid containing exogenous lipooligosaccharide synthesis genes (lsg) from Haemophilus influenzae. Analysis of these novel oligosaccharide-LPS chimeras allowed characterization of the carbohydrate structures generated by several putative glycosyltransferase genes within the lsg locus. Here, we adapted this strategy to construct a modular approach to study the synthetic properties of individual glycosyltransferases expressed alone and in combinations. To this end, a set of expression vectors containing one to four putative glycosyltransferase genes from the lsg locus, lsgC-F, were transformed into E. coli K12 (XL-1) which is defective in LPS O-antigen biosynthesis. This strategy relied on the inclusion of the H. influenzae gene product lsgG in every plasmid construct, which partially rescues the E. coli LPS biosynthesis defect by priming UDP-undecaprenyl in the WecA-dependent O-antigen synthetic pathway with N-acetyl-glucosamine (GlcNAc). This GlcNAc-undecaprenyl then served as an acceptor substrate for further carbohydrate extension by transformed glycosyltransferases. The resultant LPS-linked chimeric glycans were isolated from their E. coli constructs and characterized by mass spectrometry, methylation analysis and ELISA. These structural data allowed the specificity of various glycosyltransferases to be unambiguously assigned to individual genes. LsgF was found to transfer a galactose (Gal) to terminal GlcNAc. LsgE was found to transfer GlcNAc to Gal-GlcNAc, and both LsgF and LsgD were found to transfer Gal to GlcNAc-Gal-GlcNAc, but with differing linkage specificities. This method can be generalized and readily adapted to study the substrate specificity of other putative or uncharacterized glycosyltransferases.

  Escherichia coli, mass spectrometry, glycosyltransferase, assay, E. coli, chimera

  NCBI PubMed ID: 20208062
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: bgibson@buckinstitute.org
  Institutions: Department of Pharmaceutical Chemistry and Pharmaceutical Sciences, University of California, San Francisco, CA, USA
  Methods: HF solvolysis, de-O-acylation, SDS-PAGE, ELISA, MALDI-TOF MS, genetic methods
  
   
  
  Escherichia coli K12 (pGEM)
  CSDB ID 25295 (all data & tools)

Show legend Show as text

Structure type: structural motif or average structure
Trivial name: ceramide phosphoinositol glycan core (CPI-GC)
Compound class: LPG
Contained glycoepitopes: IEDB_135813,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_145003,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_190606,SB_173,SB_7

• Article ID: 5942
  Heiss C, Wang Z, Azadi P, Fichrova RN, Singh BN "Novel structural features of the immunocompetent ceramide phospho-inositol glycan core from Trichomonas vaginalis" - Carbohydrate Research 419 (2016) 51-59
  

  The ceramide phosphoinositol glycan core (CPI-GC) of the lipophosphoglycan of Trichomonas vaginalis is a major virulent factor of this common genitourinary parasite. While its carbohydrate composition has been reported before, its structure has remained largely unknown. We isolated the glycan portions of CPI-GC by nitrous acid deamination and hydrofluoric acid treatment and investigated their structures by methylation analysis and 1- and 2-D NMR. We found that the α-anomer of galactose is a major constituent of CPI-GC. The β-anomer was found exclusively at the non-reducing end of CPI-GC side chains. Furthermore the data showed that the rhamnan backbone is more complex than previously thought and that the inositol residue at the reducing end is linked to a 4-linked α-glucuronic acid (GlcA) residue. This appears to be the most striking and novel feature of this GPI-anchor type molecule.

  NMR, glycoconjugates, Phospho-inositol glycan core, Trichomonas vaginalis

  NCBI PubMed ID: 26671321
  Publication DOI: 10.1016/j.carres.2015.11.001
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: singhb@upstate.edu; cheiss@uga.edu
  Institutions: Complex Carbohydrate Research Center, The University of Georgia, 315 Riverbend Road, Athens, GA 30602, USA, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115, USA, Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 E Adams St, Syracuse, NY 13210, USA
  Methods: 13C NMR, 1H NMR, NMR-2D, methylation, GC-MS, HF treatment, nitrous deamination, enzymatic treatment, HPAE-PAD HPLC
  
   
  
  Trichomonas vaginalis UR1
  CSDB ID 4543 (all data & tools)

Show legend Show as text

Structure type: structural motif or average structure
Trivial name: ceramide phosphoinositol glycan core (CPI-GC)
Compound class: LPG
Contained glycoepitopes: IEDB_114701,IEDB_115136,IEDB_127513,IEDB_131174,IEDB_133754,IEDB_135610,IEDB_135813,IEDB_136105,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_140630,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_143254,IEDB_144993,IEDB_144998,IEDB_145003,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_167070,IEDB_167188,IEDB_174332,IEDB_190606,IEDB_225177,IEDB_885823,IEDB_983931,SB_173,SB_192,SB_7

• Article ID: 5942
  Heiss C, Wang Z, Azadi P, Fichrova RN, Singh BN "Novel structural features of the immunocompetent ceramide phospho-inositol glycan core from Trichomonas vaginalis" - Carbohydrate Research 419 (2016) 51-59
  

  The ceramide phosphoinositol glycan core (CPI-GC) of the lipophosphoglycan of Trichomonas vaginalis is a major virulent factor of this common genitourinary parasite. While its carbohydrate composition has been reported before, its structure has remained largely unknown. We isolated the glycan portions of CPI-GC by nitrous acid deamination and hydrofluoric acid treatment and investigated their structures by methylation analysis and 1- and 2-D NMR. We found that the α-anomer of galactose is a major constituent of CPI-GC. The β-anomer was found exclusively at the non-reducing end of CPI-GC side chains. Furthermore the data showed that the rhamnan backbone is more complex than previously thought and that the inositol residue at the reducing end is linked to a 4-linked α-glucuronic acid (GlcA) residue. This appears to be the most striking and novel feature of this GPI-anchor type molecule.

  NMR, glycoconjugates, Phospho-inositol glycan core, Trichomonas vaginalis

  NCBI PubMed ID: 26671321
  Publication DOI: 10.1016/j.carres.2015.11.001
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: singhb@upstate.edu; cheiss@uga.edu
  Institutions: Complex Carbohydrate Research Center, The University of Georgia, 315 Riverbend Road, Athens, GA 30602, USA, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115, USA, Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 E Adams St, Syracuse, NY 13210, USA
  Methods: 13C NMR, 1H NMR, NMR-2D, methylation, GC-MS, HF treatment, nitrous deamination, enzymatic treatment, HPAE-PAD HPLC
  
   
  
  Trichomonas vaginalis UR1
  CSDB ID 4551 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130646,IEDB_130651,IEDB_130654,IEDB_130701,IEDB_135813,IEDB_136044,IEDB_136045,IEDB_136104,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137485,IEDB_140108,IEDB_140116,IEDB_140122,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_141829,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_144987,IEDB_144991,IEDB_145003,IEDB_145668,IEDB_145669,IEDB_146104,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_149557,IEDB_150092,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_167070,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_190606,IEDB_221845,IEDB_423128,IEDB_461720,IEDB_540672,IEDB_548907,IEDB_742247,IEDB_983930,SB_136,SB_157,SB_165,SB_166,SB_177,SB_187,SB_195,SB_196,SB_197,SB_198,SB_30,SB_31,SB_33,SB_44,SB_62,SB_67,SB_7,SB_72,SB_73,SB_74,SB_85,SB_86,SB_88

• Article ID: 10375
  van Huystee RB, Sesto PA, O'Donell JP "Number and size of oligosaccharides linked to peanut peroxidases" - Plant Physiology and Biochemistry 30 (1992) 147-152
  
  Journal NLM ID: 9882449
  Publisher: Elsevier Science for Société Française De Physiologie Végétale
  
   
  
  Arachis hypogaea
  CSDB ID 147119 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130646,IEDB_130701,IEDB_135813,IEDB_136044,IEDB_136045,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137485,IEDB_1391962,IEDB_140108,IEDB_140116,IEDB_140122,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142489,IEDB_143794,IEDB_144562,IEDB_144983,IEDB_145003,IEDB_145668,IEDB_145669,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_150092,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_167070,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_190606,IEDB_548907,IEDB_983930,SB_137,SB_165,SB_166,SB_187,SB_195,SB_197,SB_198,SB_29,SB_30,SB_33,SB_44,SB_67,SB_7,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86,SB_88

• Article ID: 10375
  van Huystee RB, Sesto PA, O'Donell JP "Number and size of oligosaccharides linked to peanut peroxidases" - Plant Physiology and Biochemistry 30 (1992) 147-152
  
  Journal NLM ID: 9882449
  Publisher: Elsevier Science for Société Française De Physiologie Végétale
  
   
  
  Arachis hypogaea
  CSDB ID 147121 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: (->4) Asn-X-Ser/Thr (major cationic peanut peroxidase)
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_115015,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130646,IEDB_130701,IEDB_135813,IEDB_136044,IEDB_136045,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137485,IEDB_1391962,IEDB_140108,IEDB_140116,IEDB_140122,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_142489,IEDB_143794,IEDB_144562,IEDB_144983,IEDB_145003,IEDB_145668,IEDB_145669,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_149135,IEDB_150092,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_167070,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_190606,IEDB_548907,IEDB_983930,SB_137,SB_165,SB_166,SB_187,SB_195,SB_197,SB_198,SB_29,SB_30,SB_33,SB_44,SB_67,SB_7,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86,SB_88

• Article ID: 11040
  van Huystee RB, McManus MT "Glycans of higher plant peroxidases: recent observations and future speculations" - Glycoconjugate Journal 15(2) (1998) 101-106
  

  Plant peroxidases are composed of a peptide and associated heme, calcium and glycans. The 3D structure of the major cationic peanut peroxidase has revealed the sites of the heme and calcium. But the diffraction of the glycans was not sufficient to show their structure. This review presents research that has been executed to obtain putative glycans and their binding sites, and to gain an indirect insight into these glycans. It also offers approaches that will be used to determine the function of the glycans on the peanut peroxidase. Some comparisons are made with other plant glycoproteins including peroxidases from plants other than peanut.

  glycans, plant peroxidases

  NCBI PubMed ID: 9557869
  Publication DOI: 10.1023/a:1006955903531
  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Institutions: Department of Plant Sciences, The University of Western Ontario London, Ontario, Canada, Department of Plant Biology and Biotechnology, Massey University, Palmerston North, New Zealand
  
   
  
  Arachis hypogaea
  CSDB ID 61921 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: (->4) Asn-X-Ser/Thr (major cationic peanut peroxidase)
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_115015,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130646,IEDB_130651,IEDB_130654,IEDB_130701,IEDB_135813,IEDB_136044,IEDB_136045,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137485,IEDB_140108,IEDB_140116,IEDB_140122,IEDB_141793,IEDB_141794,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_144987,IEDB_144991,IEDB_145003,IEDB_145668,IEDB_145669,IEDB_146104,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_149135,IEDB_149557,IEDB_150092,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_167070,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_190606,IEDB_221845,IEDB_461720,IEDB_548907,IEDB_742247,IEDB_983930,SB_157,SB_165,SB_166,SB_177,SB_187,SB_195,SB_197,SB_198,SB_30,SB_31,SB_33,SB_44,SB_62,SB_67,SB_7,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86,SB_88

• Article ID: 11040
  van Huystee RB, McManus MT "Glycans of higher plant peroxidases: recent observations and future speculations" - Glycoconjugate Journal 15(2) (1998) 101-106
  

  Plant peroxidases are composed of a peptide and associated heme, calcium and glycans. The 3D structure of the major cationic peanut peroxidase has revealed the sites of the heme and calcium. But the diffraction of the glycans was not sufficient to show their structure. This review presents research that has been executed to obtain putative glycans and their binding sites, and to gain an indirect insight into these glycans. It also offers approaches that will be used to determine the function of the glycans on the peanut peroxidase. Some comparisons are made with other plant glycoproteins including peroxidases from plants other than peanut.

  glycans, plant peroxidases

  NCBI PubMed ID: 9557869
  Publication DOI: 10.1023/a:1006955903531
  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Institutions: Department of Plant Sciences, The University of Western Ontario London, Ontario, Canada, Department of Plant Biology and Biotechnology, Massey University, Palmerston North, New Zealand
  
   
  
  Arachis hypogaea
  CSDB ID 61357 (all data & tools)