Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide with O-unit
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_130701,IEDB_133751,IEDB_135513,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144983,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_152206,IEDB_153307,IEDB_190606,IEDB_2189047,IEDB_225177,IEDB_226811,IEDB_885823,IEDB_983930,IEDB_983931,SB_192,SB_44,SB_67,SB_7,SB_72

• Article ID: 265
  Inagaki M, Kawaura T, Wakashima H, Kato M, Nishikawa S, Kashimura N "Different contributions of the outer and inner R-core residues of lipopolysaccharide to the recognition by spike H and G proteins of bacteriophage phiX174" - FEMS Microbiology Letters 226(2) (2003) 221-227
  

  The binding of spike H and G proteins of bacteriophage phiX174 with lipopolysaccharides (LPSs) were evaluated by a competitive enzyme-linked plate assay using the biotin-labeled LPS of Escherichia coli C, one of a host strain, and the non-labeled LPSs having different R-core polysaccharide lengths. H protein promptly decreased its affinity when some saccharide residues were truncated from the outer R-core. However, G protein showed significant affinity to the LPSs lacking all the residues of the outer R-core and some of the inner R-core. Thus, G protein rather than H protein well recognized the residues of the inner R-core of LPS

  LPS, core, bacteriophage, proteins, G-protein, phiX174, host recognition, spike protein

  NCBI PubMed ID: 14553915
  Journal NLM ID: 7705721
  Publisher: Blackwell Publishing
  Correspondence: inagaki@bio.mie-u.ac.jp
  Institutions: Department of Life Science, Faculty of Bioresources, Mie University, 1515 Kamihama, Tsu, 514-8507, Mie, Japan
  Methods: DOC-PAGE, enzyme-linked plate assay
  
   
  
  Salmonella enterica Typhimurium
  CSDB ID 7141 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide with O-unit
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 265
  Inagaki M, Kawaura T, Wakashima H, Kato M, Nishikawa S, Kashimura N "Different contributions of the outer and inner R-core residues of lipopolysaccharide to the recognition by spike H and G proteins of bacteriophage phiX174" - FEMS Microbiology Letters 226(2) (2003) 221-227
  

  The binding of spike H and G proteins of bacteriophage phiX174 with lipopolysaccharides (LPSs) were evaluated by a competitive enzyme-linked plate assay using the biotin-labeled LPS of Escherichia coli C, one of a host strain, and the non-labeled LPSs having different R-core polysaccharide lengths. H protein promptly decreased its affinity when some saccharide residues were truncated from the outer R-core. However, G protein showed significant affinity to the LPSs lacking all the residues of the outer R-core and some of the inner R-core. Thus, G protein rather than H protein well recognized the residues of the inner R-core of LPS

  LPS, core, bacteriophage, proteins, G-protein, phiX174, host recognition, spike protein

  NCBI PubMed ID: 14553915
  Journal NLM ID: 7705721
  Publisher: Blackwell Publishing
  Correspondence: inagaki@bio.mie-u.ac.jp
  Institutions: Department of Life Science, Faculty of Bioresources, Mie University, 1515 Kamihama, Tsu, 514-8507, Mie, Japan
  Methods: DOC-PAGE, enzyme-linked plate assay
  
   
  
  Salmonella enterica Typhimurium TV119
  CSDB ID 7208 (all data & tools)

Show legend Show as text

Structure type: oligomer
Trivial name: Ra core oligosaccharide
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 313
  Lugowski C, Jachymek W, Niedziela T, Rowinski S "Serological characterisation of anti-endotoxin sera directed against the conjugates of oligosaccharide core of Escherichia coli type R1, R2, R3, J5 and Salmonella Ra with tetanus toxoid" - FEMS Immunology and Medical Microbiology 16 (1996) 21-30
  

  The covalent conjugates of oligosaccharide core: Escherichia coli type R1, R2, R3, J5 and Salmonella Ra with tetanus toxoid have been prepared using reaction of reductive amination. The neoglycoconjugates were good immunogens in rabbits yielding a high level of anti-lipopolysaccharide antibodies of IgG class. The antibodies were used to examine the possibility of their reactions with smooth lipopolysaccharides. We have found that all antisera were able to react with the lipopolysaccharide molecules of identical or related core type, possessing core oligosaccharides substituted with O-specific chains. These reactions were shown in both the ELISA assay and the immunoblotting test.

  oligosaccharide, core, Escherichia coli, endotoxin, serological, Salmonella, anti-endotoxin, conjugate vaccine, tetanus toxoid

  NCBI PubMed ID: 8954349
  Journal NLM ID: 9315554
  Publisher: Elsevier
  Correspondence: lugowski@immuno.pan.wroc.pl
  Institutions: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
  
   
  
  Salmonella enterica
  CSDB ID 7536 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130656,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_135813,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 2449
  Kastowsky M, Gutberlet T, Bradaczek H "Comparison of X-ray powder-diffraction data of various bacterial lipopolysaccharide structures with theoretical model conformations" - European Journal of Biochemistry 217 (1993) 771-779
  

  X-ray powder-diffraction experiments have been performed on dry samples of lipid A and various rough-mutant lipopolysaccharides (LPS) of Salmonella minnesota, Salmonella typhimurium and Escherichia coli. The diffraction patterns obtained indicated exclusively lamellar, bilayered arrangements in all samples. The periodicities were found to be in the range 4.5 nm for lipid A to 8.8 nm for Ra-LPS. Upon treatment with water-saturated air, swelling of the lamellar structures was achieved, as indicated by shifts of reflections. The increase in bilayer dimensions normally was about 0.3 nm. X-ray intensities were used for the determination of the inner bilayer structure, i.e. for calculation of the one-dimensional electron-density distribution across the bilayer. For lipid A and several Re-LPS, Rd2-LPS, Rd1-LPS and Rc-LPS samples, a striking coincidence of the electron-density distributions in the lipid-A domain was found, suggesting that in all these structures the lipid-A portion is similarly arranged. For Rb1 and Ra-LPS the lipid-A domain could not be resolved due to the limited number of observed reflections. For other Re-mutant lipopolysaccharide samples, quite different X-ray patterns were obtained. Some samples yielded diffraction patterns indicating a very high state of order in the lipid-A domain, whereas, in others, a significantly reduced order in the lipid-A domain was inferred. Comparison of the X-ray data with features of a calculated three-dimensional molecular model of lipopolysaccharide revealed reasonable agreement in molecular dimensions and bilayer structure.

  NCBI PubMed ID: 8223620
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Institutions: Institut für Kristallographie, Freien Universität Berlin, Germany
  Methods: X-ray
  
   
  
  Escherichia coli EH100, Escherichia coli R3, Salmonella minnesota R60, Salmonella typhimurium TV119
  CSDB ID 124340 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_137777,IEDB_137778,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 2632
  Wollin R, Creeger ES, Rothfield LI, Stocker BAD, Lindberg AA "Salmonella typhimurium mutants defective in UDP-D-galactose: Lipopolysaccharide a1,6-D-galactosyltransferase. Structural, immunochemical, and enzymologic studies of rfaB mutants" - Journal of Biological Chemistry 258 (1983) 3769-3774
  
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Methods: 1H NMR, GC-MS
  
   
  
  Salmonella typhimurium rfaB
  CSDB ID 133996 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: LPS
Contained glycoepitopes: IEDB_130656,IEDB_130670,IEDB_130693,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 2657
  Luk JMC, Lind SM, Tsang RSW, Lindberg AA "Epitope mapping of four monoclonal antibodies recognizing the hexose core domain of Salmonella lipopolysaccharide" - Journal of Biological Chemistry 266 (1991) 23215-23225
  
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  
   
  
  Salmonella typhimurium SH 4809
  CSDB ID 136230 (all data & tools)
• Article ID: 2728
  Jansson PE, Wollin R, Bruse GW, Lindberg AA "The conformation of core oligosaccharides from Escherichia coli and Salmonella typhimurium lipopolysaccharides as predicted by semi-empirical calculations" - Journal of Molecular Recognition 2 (1989) 25-36
  
  Journal NLM ID: 9004580
  Publisher: Chichester, Sussex, UK: John Wiley & Sons
  Methods: conformation analysis
  
   
  
  Escherichia coli, Salmonella typhimurium
  CSDB ID 140665 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 2730
  Seydel U, Koch MHJ, Brandenburg K "Structural polymorphisms of rough mutant lipopolysaccharides Rd to Ra from Salmonella minnesota" - Journal of Structural Biology 110 (1993) 232-243
  

  The structural polymorphisms of rough mutant lipopolysaccharides (LPS) Rd, Rc, Rb, and Ra from Salmonella minnesota (strains R4, R7, Rz, R5, R345, and R60, respectively) were investigated as a function of temperature, water content, and Mg2+ concentration. The gel to liquid crystalline (beta<==>alpha) phase transition temperature (Tc) and the state of order within each phase were measured by Fourier transform infrared spectroscopy. The amount of bound water was determined by differential scanning calorimetry and the three-dimensional structures in each phase state were characterized by synchrotron radiation X-ray diffraction. The results indicate an extremely complex dependence of the structural behavior of LPS on ambient conditions. The beta<==>alpha acyl chain melting temperatures at high water contents (95-97%), Tc = 31 to 32 degrees C for LPS Rd, 33 to 35 degrees C for LPS Rc to Rb, and 36 degrees C for LPS Ra, increase with decreasing water content and in the presence of Mg2+ cations with a concomitant broadening of the transition range. Below 30 to 50% water content, no distinct phase transitions can be observed. These effects are most pronounced for LPS with the shortest sugar chains. Below 50% water content, only lamellar structures can be observed in the temperature range 5 to 80 degrees C, independent of the Mg2+ concentration. Above 50% water concentration, for large [LPS]:[Mg2+] molar ratios the predominant structure above Tc is nonlamellar; for smaller [LPS]:[Mg2+] molar ratios a superposition of lamellar and nonlamellar structures is found. For all LPS Rd to Rb at low Mg2+ concentrations, the phase transition is connected with a change in the three-dimensional structure from lamellar or mixed lamellar/nonlamellar to a pure nonlamellar, probably cubic structure. The tendency to form non-lamellar structures decreases with the length of the core oligosaccharide. At an equimolar ratio of [LPS] and [Mg2+] a multibilayered organization is observed. Some of the nonlamellar structures are cubic phases with periodicities between 12 and 16 nm. The molecular dimensions of the single endotoxin molecules in the absence and the presence of external water are estimated from the different lamellar periodicities, including those of free lipid A and deep rough mutant LPS Re. These observations are discussed with respect to the biological importance of LPS as a potent inducer of biological effects in mammals.

  NCBI PubMed ID: 8373704
  Publication DOI: 10.1006/jsbi.1993.1026
  Journal NLM ID: 9011206
  Institutions: Forzchungsinstitut Borstel, D-23845 Borstel, Germany, European Molecular Biology Laboratory, c/o DESY, D-22607 Hamburg 52, Germany
  
   
  
  Salmonella minnesota R60
  CSDB ID 141925 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: (2->6) lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 3272
  Rietschel ET, Kirikae T, Schade FU, Mamat U, Schmidt G, Loppnow H, Ulmer AJ, Zähringer U, Seydel U, Di Padova F, Schreier M, Brade H "Bacterial endotoxin: Molecular relationships of structure to activity and function" - FASEB Journal 8 (1994) 217-225
  

  Endotoxins of Gram-negative microbes fulfill as components of the outer membrane a vital function for bacterial viability and, if set free, induce inmammalians potent pathophysiological effects. Chemically, they are lipopolysaccharides (LPS) consisting of an 0-specific chain, a core oligosaccharide, and a lipid component, termed lipid A. The latter determines the endotoxicactivities and, together with the core constituent Kdo, essential functions for bacteria. The primary structure of lipid A of various bacterial origin has been elucidated and lipid A of Escherichia coli has been chemically synthesized. The biological analysis of synthetic lipid A partial structures proved that the expression of endotoxic activity depends on a unique primary structure and a peculiar endotoxic conformation. The biological lipid A effects are mediated by macrophage-derived bioactive peptides such as tumor necrosis factor a (TNF). Macrophages possess LPS receptors, and the lipid A regions involved in specific binding and cell activation have been characterized. Synthetic lipid A partial structures compete the specific binding of LPS or lipid A and antagonistically inhibit the production of LPS-induced TNF. LPS toxicity, in general, and the ability of LPS to induce TNF are also suppressed by a recently developed monoclonal antibody (IgG2a), which is directed against an epitope located in the core oligosaccharide. At present we determine molecular and submolecular details of the specificity of the interaction of lipid A with responsive host cells with the ultimate aim to provide pharmacological or immunological therapeutics that reduce or abolish the fatal inflammatory consequences of endotoxicosis.

  Lipopolysaccharide, lipid A, endotoxin, activity, function, Gram-negative, tumor necrosis factor, interleukin 1, anti-LPS antibodies, LPS antagonists - monocytes, sphingogiycoiipid

  NCBI PubMed ID: 8119492
  Journal NLM ID: 8804484
  Publisher: Bethesda, MD: Federation of American Societies for Experimental Biology
  Institutions: Forschungsinstitut Borstel, Institut für Experimentelle Biologie und Medizin, Germany
  Methods: biological assays
  
   
  
  Salmonella
  CSDB ID 22446 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: Lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 3280
  Oscarson S, Ritzén H "Synthesis of a hexasaccharide corresponding to part of the heptose-hexose region of the Salmonella Ra core, and a penta- and a tetra-saccharide that compose parts of this structure" - Carbohydrate Research 254 (1994) 81-90
  

  The synthesis of the hexasaccharide 2-(4-trifluoroacetamidophenyl)ethyl O-α-D-galactopyranosyl-(1→3)-[O-α-D-galactopyranosyl-(1→6)]- O-α-D-glucopyranosyl-(1→3)-[O-L-glycero-α-D-manno-heptopyranosyl-(1→7)]-O-L-glycero-α-D-manno-heptopyranosyl-(1→3)-L-glycero-α-D-manno-heptopyranoside, corresponding to the heptose and part of the hexose region in the Salmonella Ra core, is described. Syntheses of the pentasaccharide 2-(4-trifluoroacetamidophenyl)ethyl O-α-D-galactopyranosyl-(1→3)-O-α-D-glucopyranosyl-(1→3)-[O-L-glycero-α-D-manno-heptopyranosyl-(1→7)]-O-L-glycero-α-D-manno- heptopyranosyl-(1→3)-L-glycero-α-D-manno-heptopyranoside and the tetrasaccharide 2-(4-trifluoroacetamidophenyl)ethyl O-α-D-glucopyranosyl-(1→3)-[O-L-glycero-α-D-manno-heptopyranosyl-(1→7)]-O-L-glycero-α-D-manno-heptopyranosyl-(1→3)-L-glycero-α-D-manno-heptopyranoside are also described. Coupling of methyl 2,3,4,6-tetra-O-benzyl-1-thio-β-D-glucopyranoside and methyl 2-O-benzyl-4,6-O-benzylidene-3-O-(2,3,4,6-tetra-O-benzyl-α-D-galactopyranosyl)-1-thio-β-D-glucopyranoside to a triheptoside derivative with a free HO-3', using dimethyl(methylthio)sulfonium triflate and N-iodosuccinimide-silver triflate as promoters, gave the protected tetra- and penta-saccharide, respectively. Removal of the benzylidene group from the pentasaccharide followed by a regio- and stereo-selective coupling using halide-assisted conditions and 2,3,4,6-tetra-O-benzyl-α-D-galactopyranosyl bromide as donor gave the protected hexasaccharide. Deprotection then gave the target structures.

  synthesis, structure, core, heptose, tetrasaccharide, hexasaccharide, chemistry, group, Salmonella, region, derivative, hexose, pentasaccharide, stereoselective, PDF, methyl, free, removal, Ra core

  NCBI PubMed ID: 8180998
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Sweden.
  Methods: 13C NMR, 1H NMR, chemical synthesis
  
   
  
  Salmonella
  CSDB ID 22410 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Trivial name: core region
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 3622
  Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. II. The structure of the core region" - Biochemistry (Moscow) 58(2) (1993) 84-99
  

  This review summarizes data on the structure of the core of bacterial lipopolysaccharides (LPS), an oligosaccharide which binds the lipid moiety of LPS to the O-antigenic polysaccharide chain. Both S-strains with complete LPS and R-mutants having various defects of core biosynthesis are considered. The role of the core in the functioning of the outer membrane and in the manifestation of antigenic specificity of LPS is discussed.

  Lipopolysaccharide, antigen, lipopolysaccharides, LPS, structure, core, bacteria, core region, region, Gram-negative bacteria, gram negative bacteria, Gram-negative, review, outer membrane, bacterial lipopolysaccharide

  Journal NLM ID: 0376536
  Publisher: Nauka/Interperiodica
  Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow (Russian Federation)
  
   
  
  Salmonella
  CSDB ID 24950 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_150901,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 4317
  Huang JX, Azad MA, Yuriev E, Baker MA, Nation RL, Li J, Cooper MA, Velkov T "Molecular Characterization of Lipopolysaccharide Binding to Human a-1-Acid Glycoprotein" - Journal of Lipids 2012 (2012) 475153
  

  The ability of AGP to bind circulating lipopolysaccharide (LPS) in plasma is believed to help reduce the proinflammatory effect of bacterial lipid A molecules. Here, for the first time we have characterized human AGP binding characteristics of the LPS from a number of pathogenic Gram-negative bacteria: Escherichia coli, Salmonella typhimurium, Klebsiella pneumonia, Pseudomonas aeruginosa, and Serratia marcescens. The binding affinity and structure activity relationships (SAR) of the AGP-LPS interactions were characterized by surface plasma resonance (SPR). In order to dissect the contribution of the lipid A, core oligosaccharide and O-antigen polysaccharide components of LPS, the AGP binding affinity of LPS from smooth strains, were compared to lipid A, Kdo2-lipid A, R(a), R(d), and R(e) rough LPS mutants. The SAR analysis enabled by the binding data suggested that, in addition to the important role played by the lipid A and core components of LPS, it is predominately the unique species- and strain-specific carbohydrate structure of the O-antigen polysaccharide that largely determines the binding affinity for AGP. Together, these data are consistent with the role of AGP in the binding and transport of LPS in plasma during acute-phase inflammatory responses to invading Gram-negative bacteria.

  O-antigen, lipid A, gram negative bacteria, lipopolysaccharide-binding, AGP binding, SPR

  NCBI PubMed ID: 23316371
  Publication DOI: 10.1155/2012/475153
  Journal NLM ID: 101553819
  Publisher: Cairo: Hindawi Pub Corp
  Correspondence: Tony Velkov
  Institutions: Institute for Molecular Bioscience, The University of Queensland, 306 Carmody Road, St. Lucia, QLD 4072, Australia, Drug Development and Innovation, Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC 3052, Australia, Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC 3052, Australia, Priority Research Centre in Reproductive Science, School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia
  Methods: molecular modeling, fluorescence spectroscopy, surface plasmon resonance (SPR)
  
   
  
  Salmonella typhimurium
  CSDB ID 28408 (all data & tools)
  Salmonella typhimurium TV119
  CSDB ID 28409 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130656,IEDB_130659,IEDB_130670,IEDB_130693,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 4368
  Patel KB, Toh E, Fernandez XB, Hanuszkiewicz A, Hardy GG, Brun YV, Bernards MA, Valvano MA "Functional characterization of UDP-Glucose:Undecaprenyl-Phosphate Glucose-1-Phosphate Transferases of Escherichia coli and Caulobacter crescentus" - Journal of Bacteriology 194(10) (2012) 2646-2657
  

  Escherichia coli K-12 WcaJ and Caulobacter crescentus HfsE, PssY and PssZ enzymes are predicted to initiate the synthesis of colanic acid (CA) capsule and holdfast polysaccharide, respectively. These proteins belong to a prokaryotic family of membrane enzymes that catalyze the formation of a phosphoanhydride bond joining a hexose-1-phosphate with undecaprenyl phosphate (Und-P). Here, in vivo complementation assays of an E. coli K-12 wcaJ mutant demonstrate that WcaJ and PssY can complement CA synthesis. Furthermore, WcaJ can restore holdfast production in C. crescentus. In vitro transferase assays demonstrated that both WcaJ and PssY utilize UDP-glucose but not UDP-galactose. However, in a strain of Salmonella enterica Typhimurium deficient in the WbaP O antigen initiating galactosyltransferase, complementation with WcaJ or PssY resulted in O antigen production. GC-MS analysis of the LPS revealed the attachment of both CA and O antigen molecules to lipid A-core OS. Therefore, while UDP-glucose is the preferred substrate of WcaJ and PssY, these enzymes can also utilize UDP-galactose. This unexpected feature of WcaJ and PssY may help map specific residues responsible for the nucleotide diphosphate specificity in these or similar enzymes. Also, the reconstitution of O antigen synthesis in Salmonella, CA capsule in E. coli and holdfast provide biological assays of high sensitivity to examine the sugar-1-phosphate transferase specificity of heterologous proteins.

  O-antigen, specificity, Salmonella enterica, galactosyltransferase, colanic acid, Typhimurium, UDP-glucose, Escherichia coli K12, Caulobacter crescentus

  NCBI PubMed ID: 22408159
  Publication DOI: 10.1128/JB.06052-11
  Journal NLM ID: 2985120R
  Publisher: American Society for Microbiology
  Correspondence: Miguel A. Valvano
  Institutions: Departments of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada
  Methods: DNA sequencing, GC-MS, SDS-PAGE, glycosyltransferase assays, genetic methods
  
   
  
  Salmonella enterica sv. Typhimurium
  CSDB ID 28557 (all data & tools)

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Structure type: oligomer
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130656,IEDB_130670,IEDB_130693,IEDB_133751,IEDB_136906,IEDB_137472,IEDB_140088,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

• Article ID: 5145
  De Benedetto G, Salvini L, Gotta S, Cescutti P, Micoli F "Investigation on Sugar-Protein Connectivity in Salmonella O-Antigen Glycoconjugate Vaccines" - Bioconjugate Chemistry 29(5) (2018) 1736-1747
  

  Invasive nontyphoidal Salmonella disease, for which licensed vaccines are not available, is a leading cause of bloodstream infections in Africa. The O-antigen portion of lipopolysaccharide is a good target for protective immunity. Covalent conjugation of the O-antigen to a carrier protein increases its immunogenicity and O-antigen based glycoconjugate vaccines are currently under investigation at the preclinical stage. We developed a conjugation chemistry for linking O-antigen to CRM197 carrier protein, through sequential insertion of adipic acid dihydrazide (ADH) and adipic acid bis( N-hydroxysuccinimide) ester (SIDEA) as linkers, without impacting O-antigen chain epitopes. Here the resulting sugar-protein connectivity has been investigated in detail. The core portion of the lipopolysaccharide was used as a model molecule to prepare CRM197 conjugates, making structural investigations easier. The first step of reductive amination with ADH involves the terminal 3-deoxy-d- manno-oct-2-ulosonic acid (KDO) residue of the core region. The second reaction step resulted not to be selective, as SIDEA reacted with both ADH and pyrophosphorylethanolamine (PPEtN) of the core region, independently from the pH at which the reaction was performed. Peptide mapping analysis of the deglycosylated core-CRM197 conjugates confirmed that lysine residues of CRM197 were linked to SIDEA not only through KDO-ADH but also through PPEtN. This analysis also confirmed that the conjugation chemistry is random on the protein, involving a large number of lysine residues, particularly the surface exposed ones. The method for core-CRM197 characterization was successfully extended to O-antigen-CRM197 conjugate, confirming the results obtained with the core. This study not only allowed full characterization of OAg-CRM197 conjugates, but can be applied to optimize synthesis and characterization of other OAg-based glycoconjugate vaccines. Analytical methods to investigate saccharide-protein connectivity are also of fundamental importance to study the relationship between glycoconjugate structure and immune response induced.

  Lipopolysaccharide, synthesis, disease, O-antigen, Kdo, epitopes, Salmonella, core region, vaccines, immunogenicity, conjugate, model, CRM197, glycoconjugate vaccine, Peptide Mapping

  NCBI PubMed ID: 29697244
  Publication DOI: 10.1021/acs.bioconjchem.8b00178
  Journal NLM ID: 9010319
  Publisher: Washington, DC: American Chemical Society
  Correspondence: francesca.x.micoli@gsk.com
  Institutions: Fondazione Toscana Life Sciences , via Fiorentina 1, 53100 Siena, Italy, Dipartimento di Scienze della Vita, Universita degli Studi di Trieste, Ed. C11, via L. Giorgieri 1, 34127 Trieste, Italy, GSK Vaccines Institute for Global Health (GVGH) S.r.l., via Fiorentina 1, 53100 Siena, Italy
  Methods: gel filtration, SDS-PAGE, mild acid hydrolysis, MALDI-MS, HPAEC-PAD, RP-HPLC, conjugation, LC-ESI-MS/MS, HPLC-SEC, HR-ESI-MS
  
   
  
  Salmonella enterica sv. Typhimurium 1418 ΔtolR ΔwbaP
  CSDB ID 12890 (all data & tools)