Show legend Show as text

Structure type: oligomer
Trivial name: core-lipid A backbone, epitope J
Compound class: core oligosaccharide, LPS
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135394,IEDB_137340,IEDB_140092,IEDB_141807,IEDB_150755,IEDB_150756,IEDB_150760,IEDB_150901,IEDB_150908,IEDB_151531,IEDB_151769

• Article ID: 814
  Holst O, Bock K, Brade L, Brade H "The structures of oligosaccharide bisphosphates isolated from the lipopolysaccharide of a recombinant Escherichia coli strain expressing the gene gseA [3-deoxy-D-manno-octulopyranosonic acid (Kdo) transferase] of Chlamydia psittaci 6BC" - European Journal of Biochemistry 229 (1995) 194-200
  

  The lipopolysaccharide from the recombinant strain Escherichia coli F515-140 containing the cloned gene gseA [3-deoxy-D-manno-octulopyranosonic acid (Kdo) transferase] from Chlamydia psittaci 6BC was isolated and sequentially de-O-acylated and de-N-acylated. The products were separated by high-performance anion-exchange chromatography into three fractions, two of which contained a single compound. Their structures were elucidated by high-field NMR spectroscopy as α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcN-(1→6)-α-D-GlcN 1,4'-P2 (compound 1) (tetrasaccharide bisphosphate) [Holst, O., Broer, W., Thomas-Oates, J. E., Mamat, U. & Brade, H. (1993) Eur. J. Biochem. 214, 703-710] and α-Kdo-(2→4)-[α-Kdo-(2→8)-]-α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcN-(1→6)-α-D-GlcN 1,4'-P2 (compound 4) (hexasaccharide bisphosphate). The third fraction comprised two pentasaccharide bisphosphates, which could be separated by affinity chromatography using an immobilized monoclonal antibody specific for the trisaccharide α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo. The bound fraction was identified as α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcN-(1→6)-α-D-GlcN 1,4'-P2 (compound 2) [Holst, O., Broer, W., Thomas-Oates, J. E., Mamat, U. & Brade, H. (1993) Eur. J. Biochem. 214, 703-710], whereas the unbound fraction was identified as α-Kdo-(2→4)-α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcN-(1→6)-α-D-GlcN 1,4'-P2 (compound 3). This novel Kdo tetrasaccharide extends our knowledge on multifunctional Kdo transferases.

  Lipopolysaccharide, LPS, oligosaccharide, structure, core, gene, strain, Escherichia, Escherichia coli, acid, Kdo, transferase, phosphate, Chlamydia, recombinant, bisphosphate, Chlamydia psittaci, gseA, HPLC

  NCBI PubMed ID: 7744029
  Publication DOI: 10.1111/j.1432-1033.1995.0194l.x
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Correspondence: oholst@fz-borstel.de
  Institutions: Division of Biochemical Microbiology, Forschungsinstitut Borstel, Germany, Division of Biochemical Microbiology, Institut fur Experimentelle Biologie und Medizin, Forschungsinstitut Borstel, Germany, Department of Chemistry, Carisberg Laboratory, Copenhagen, Denmark
  Methods: 13C NMR, 1H NMR, GLC-MS, 31P NMR, HPAEC, affinity chromatography
  
   
  
  Escherichia coli F515-140
  CSDB ID 2503 (all data & tools)
• Article ID: 1157
  Rund S, Lindner B, Brade H, Holst O "Structural analysis of the lipopolysaccharide from Chlamydophila psittaci strain 6BC" - European Journal of Biochemistry 267(18) (2000) 5717-5726
  

  The lipopolysaccaride of Chlamydophila psittaci 6BC was isolated from tissue culture-grown elementary bodies using a modified phenol/water procedure followed by extraction with phenol/chloroform/light petroleum. Compositional analyses indicated the presence of 3-deoxy-Dmanno-oct-2-ulosonic acid, GlcN, organic bound phosphate and fatty acids in a molar ratio of approximately 3. 3 : 2 : 1.8 : 4.6. Deacylated lipopolysaccharide was obtained after successive microscale treatment with hydrazine and potassium hydroxide, and was then separated by high performance anion-exchange chromatography into two major fractions, the structures of which were determined by 600 MHz NMR spectroscopy as α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcpN-(1→6)-α-D-GlcpN 1,4'-bisphosphate and α-Kdo-(2→4)-[α-Kdo-(2→8)]-α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcpN-(1→6)-α-D-GlcpN 1,4'-bisphosphate. The distribution of fatty acids in lipid A was determined by compositional analyses andmatrix-assisted laser desorption/ionization time-of-flight mass spectrometry experiments on lipid A and de-O-acylated lipid A. It was shown that the carbohydrate backbone of lipid A is replaced by a complex mixture of fatty acids, including long-chain and branched (R)-configured 3-hydroxy fatty acids, the latter being exclusively present in an amide linkage

  Lipopolysaccharide, strain, structural, serotype, analysis, structural analysis, Chlamydophila psittaci, NMR spectroscopy, mass spectrometry

  NCBI PubMed ID: 10971582
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Correspondence: oholst@fz-borstel.de
  Institutions: Divisions of Medical and Biochemical Microbiology and Biophysics, Research Center Borstel, Center for Medicine and Biosciences, Germany
  Methods: 13C NMR, 1H NMR, NMR-2D, MALDI-TOF MS
  
   
  
  Chlamydophila psittaci 6BC
  CSDB ID 4460 (all data & tools)
• Article ID: 1386
  Brabetz W, Lindner B, Brade H "Comparative analyses of secondary gene products of 3-deoxy-D-manno-oct-2-ulosonic acid transferases from Chlamydiaceae in Escherichia coli K-12" - European Journal of Biochemistry 267(17) (2000) 5458-5465
  

  The waaA gene encoding the essential, lipopolysaccharide (LPS)-specific 3-deoxy-Dmanno-oct-2-ulosonic acid (Kdo) transferase was inactivated in the chromosome of a heptosyltransferase I and II deficient Escherichia coli K-12 strain by insertion of gene expression cassettes encoding the waaA genes of Chlamydia trachomatis, Chlamydophila pneumoniae or Chlamydophila psittaci. The three chlamydial Kdo transferases were able to complement the knockout mutation without changing the growth or multiplication behaviour. The LPS of the mutants were serologically and structurally characterized in comparison to the LPS of the parent strain using compositional analyses, high performance anion exchange chromatography, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and specific monoclonal antibodies. The data show that chlamydial Kdo transferases can replace in E. coli K-12 the host's Kdo transferase and retain the product specificities described in their natural background. In addition, we unequivocally proved that WaaA from C. psittaci transfers predominantly four Kdo residues to lipid A, forming a branched tetrasaccharide with the structure α-Kdo-(2→8)-[α-Kdo-(2→4)]-α-Kdo-(2→4)-α-Kdo.

  Lipopolysaccharide, LPS, gene, Escherichia, Escherichia coli, 3-deoxy-D-manno-oct-2-ulosonic acid, acid, Kdo, transferase, transferases, Chlamydiaceae, genetic complementation, multifunctional 3-deoxy-D-manno-oct-2, product specificity, ulosonic acid transferases

  NCBI PubMed ID: 10951204
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Correspondence: wbrabetz@fz-borstel.de
  Institutions: Division of Medical and Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences, Borstel, Germany, Divizion of Medical and Biochemical Microbiology, Research Centre Borstel, Centre for Medicine and Biosciences,Borstel,Germany
  
   
  
  Chlamydophila psittaci 6BC, Escherichia coli K12 (WBB52)
  CSDB ID 6287 (all data & tools)
• Article ID: 3825
  Brooks CL, Müller-Loennies S, Borisova SN, Brade L, Kosma P, Hirama T, MacKenzie CR, Brade H, Evans SV "Antibodies raised against chlamydial lipopolysaccharide antigens reveal convergence in germline gene usage and differential epitope recognition" - Biochemistry 49(3) (2010) 570-581
  

  The structures of antigen-binding fragments from two related monoclonal antibodies have been determined to high resolution in the presence of several carbohydrate antigens raised against chlamydial lipopolysaccharide. With the exception of CDR H3, antibodies S54-10 and S73-2 are both derived from the same set of germline gene segments as the previously reported structures S25-2 and S45-18. Despite this similarity, the antibodies differ in specificity and the mechanism by which they recognize their cognate antigen. S54-10 uses an unrelated CDR H3 to recognize its antigen in a fashion analogous to S45-18; however, S73-2 recognizes the same antigen as S45-18 and S54-10 in a wholly unrelated manner. Together, these antibody-antigen structures provide snapshots into how the immune system uses the same set of inherited germline gene segments to generate multiple possible specificities that allow for differential recognition of epitopes and how unrelated CDR H3 sequences can result in convergent binding of clinically relevant bacterial antigens.

  antibodies, epitope, recognition, antigens, Chlamydia, binding

  NCBI PubMed ID: 20000757
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Correspondence: hebra@fz-borstel.de; svevans@uvic.ca
  Institutions: University of Victoria, Department of Biochemistry and Microbiology, PO Box 3055 STN CSC, Victoria, BC, Canada V8P 3P6
  Methods: ELISA, serological methods, genetic methods, crystallization, surface plasmon resonance (SPR)
  
   
  
  Chlamydia psittaci
  CSDB ID 25551 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_130662,IEDB_135813,IEDB_137340,IEDB_141807,IEDB_150756,IEDB_150901,IEDB_151531,IEDB_151769,IEDB_156486,IEDB_164045

• Article ID: 905
  Kosma P, Reiter A, Hofinger A, Brade L, Brade H "Synthesis of neoglycoproteins containing Kdo epitopes specific for Chlamydophila psittaci lipopolysaccharide" - Journal of Endotoxin Research 6(1) (2000) 57-69
  

  Lipopolysaccharide, synthesis, Kdo, epitope, epitopes, specific, neoglycoprotein, neoglycoproteins

  Journal NLM ID: 9433350
  Publisher: Maney Publishing
  Correspondence: pkosma@edv2.boku.ac.at
  Institutions: Department of Chemistry, University of Agricultural Sciences, Muthgasse, Vienna, Austria
  Methods: NMR
  
   
  
  Chlamydophila psittaci
  CSDB ID 3003 (all data & tools)

Show legend Show as text

Structure type: fragment of a bigger structure
Aglycon: lipid A
Trivial name: inner core
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 1440
  Gronow S, Brade H "Lipopolysaccharide biosynthesis: which steps do bacteria need to survive?" - Journal of Endotoxin Research 7(1) (2001) 3-23
  

  A detailed knowledge of LPS biosynthesis is of the utmost importance in understanding the function of the outer membrane of Gram-negative bacteria. The regulation of LPS biosynthesis affects many more compartments of the bacterial cell than the outer membrane and thus contributes to the understanding of the physiology of Gram-negative bacteria in general, on the basis of which only mechanisms of virulence and antibiotic resistance can be studied to find new targets for antibacterial treatment. The study of LPS biosynthesis is also an excellent example to demonstrate the limitations of 'genomics' and 'proteomics', since secondary gene products can be studied only by the combined tools of molecular genetics, enzymology and analytical structural biochemistry. Thus, the door to the field of 'glycomics' is opened.

  Lipopolysaccharide, biosynthesis, lipopolysaccharides, LPS, metabolism, microbiology, lipopolysaccharide biosynthesis, gram negative bacteria, membrane

  NCBI PubMed ID: 11521077
  Publication DOI: 10.1177/09680519010070010301
  Journal NLM ID: 9433350
  Publisher: Maney Publishing
  Correspondence: sgronow@fz-borstel.de
  Institutions: Division of Medical and Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences, Borstel, Germany
  
   
  
  Chlamydophila psittaci
  CSDB ID 31646 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 1459
  Holst O "Chemical structure of the core region of lipopolysaccharides - an update" - Trends in Glycoscience and Glycotechnology 14(76) (2002) 87-103
  

  Lipopolysaccharides (LPS) are the endotoxins of Gram-negative bacteria and very well known for their immunological, pharmacological and pathophysiological effects displayed in eucaryotic cells and organisms. To date, much emphasis has been put on the elucidation of the chemical structures of LPS and on their relation, or that of substructures, to the various biological effects. The lipid part of LPS, the lipid A, was proven to represent the toxic principle of endotoxin. However, lipid A toxicity depends strongly on its structure, and is influenced by a second region of LPS, the core region, that is covalently linked to lipid A. Also, the core region possesses immunogenic properties. Therefore, complete structural analyses of the core region and the comparison of its structures with biological features of LPS are of high importance for a better understanding of LPS action, and one prerequesite for strategies aimed at the treatment of endotoxicosis. In the past, quite a number of structures of the core regions from various Gram-negative bacteria were published and summarized in several overviews. The present review adds to this knowledge those structures that were published between October 1998 and December 2001.

  lipopolysaccharides, heptose, Kdo, chemical structure, core region, tructure

  Publication DOI: 10.4052/tigg.14.87
  Journal NLM ID: 9425898
  Correspondence: oholst@fz-borstel.de
  Institutions: Structural Biochemistry, Research Center Borstel, Center for Medicine and Biosciences, Borstel, Germany
  
   
  
  Chlamydophila psittaci
  CSDB ID 31818 (all data & tools)
• Article ID: 3693
  Gronow S, Lindner B, Brade H, Müller-Loennies S "Kdo-(2->8)-Kdo-(2->4)-Kdo but not Kdo-(2->4)-Kdo-(2->4)-Kdo is an acceptor for transfer of L-glycero-a-D-manno-heptose by Escherichia coli heptosyltransferase I (WaaC)" - Innate Immunity 15(1) (2009) 13-23
  

  Early steps in the biosynthesis of lipopolysaccharide (LPS) involve the transfer of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) to lipid A. Whereas Kdo transferases (WaaA) of Escherichia coli generate a (2→4)-linked Kdo disaccharide, Chlamydiae contain tri- or tetra-functional WaaA generating oligosaccharides with (2→8)- and (2→4)-linkages between Kdo. It has been suggested that the transfer of L-glycero-α-D-manno-heptose (Hep) to Kdo by an E. coli WaaC may not be possible in the presence of (2→8)-linked Kdo. E. coli double-mutants deficient in heptosyltransferases I (waaC) and II (waaF) and expressing waaA of Chlamydiae instead of their own, make Chlamydia-type Kdo oligosaccharides which are attached to an E. coli lipid A. Using such strains expressing waaA of Chlamydophila pneumoniae, Chlamydophila psittaci, or Chlamydia trachomatis, we have studied the effect of E. coli waaC gene expression on LPS structure. Structural analyses revealed the formation of two novel oligosaccharides Hep-(1→5)[Kdo-(2→4)]-Kdo and Hep-(1→5)[Kdo-(2→8)-Kdo-(2→4)]-Kdo showing that Hep is transferred in the presence of (2→8)-linked Kdo. Surprisingly, the transfer of Hep onto Kdo-(2→4)-Kdo-(2→4)-Kdo did not occur, despite the fact that Hep-(1→5)[Kdo-(2→4)-Kdo-(2→4)]-Kdo is found in nature as a partial structure of E. coli LPS. The premature end of the biosynthesis and incorporation of Hep into the LPS indicated that WaaC had access to the substrate before Kdo transfer was completed. We have observed differences between WaaA of C. trachomatis, C. pneumoniae and C. psittaci which indicate mechanistic differences between these Kdo transferases

  biosynthesis, LPS, Escherichia coli, Kdo, Hep, WaaA, WaaC, WaaF, Chlamydiae

  NCBI PubMed ID: 19201821
  Journal NLM ID: 101469670
  Publisher: Sage Publications
  Correspondence: sml@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany
  Methods: 13C NMR, 1H NMR, SDS-PAGE, 31P NMR, ESI-MS, ESI-FTICR-MS, Western blotting, composition analysis, genetic methods
  
   
  
  Chlamydia psittaci
  CSDB ID 23403 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A
Trivial name: Kdo region of LPS
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 3163
  Müller-Loennies S, Gronow S, Brade L, Mackenzie R, Kosma P, Brade H "A monoclonal antibody against a carbohydrate epitope in lipopolysaccharide differentiates Chlamydophila psittaci from Chlamydophila pecorum, Chlamydophila pneumoniae, and Chlamydia trachomatis" - Glycobiology 16(3) (2006) 184-196
  

  Lipopolysaccharide (LPS) of Chlamydophila psittaci but not of Chlamydophila pneumoniae or Chlamydia trachomatis contains a tetrasaccharide of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) of the sequence Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo. After immunization with the synthetic neoglycoconjugate antigen Kdo(2→8)[Kdo(2→4)]Kdo(2→4) Kdo-BSA, we obtained the mouse monoclonal antibody (mAb) S69-4 which was able to differentiate C. psittaci from Chlamydophila pecorum, C. pneumoniae, and C. trachomatis in double labeling experiments of infected cell monolayers and by enzyme-linked immunosorbent assay (ELISA). The epitope specificity of mAb S69-4 was determined by binding and inhibition assays using bacteria, LPS, and natural or synthetic Kdo oligosaccharides as free ligands or conjugated to BSA. The mAb bound preferentially Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo(2→4) with a K(d) of 10 microM, as determined by surface plasmon resonance (SPR) for the monovalent interaction using mAb or single chain Fv. Cross-reactivity was observed with Kdo(2→4)Kdo(2→4)Kdo but not with Kdo(2→8)Kdo(2→4)Kdo, Kdo disaccharides in 2→4- or 2→8-linkage, or Kdo monosaccharide. MAb S69-4 was able to detect LPS on thin-layer chromatography (TLC) plates in amounts of <10 ng by immunostaining. Due to the high sensitivity achieved in this assay, the antibody also detected in vitro products of cloned Kdo transferases of Chlamydia. The antibody can therefore be used in medical and veterinarian diagnostics, general microbiology, analytical biochemistry, and studies of chlamydial LPS biosynthesis. Further contribution to the general understanding of carbohydrate-binding antibodies was obtained by a comparison of the primary structure of mAb S69-4 to that of mAb S45-18 of which the crystal structure in complex with its ligand has been elucidated recently (Nguyen et al., 2003, Nat. Struct. Biol., 10, 1019-1025).

  Kdo, diagnostic, neoglycoconjugate, immunofluorescence

  NCBI PubMed ID: 16282606
  Publication DOI: 10.1093/glycob/cwj055
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany, Institute for Biological Sci-ences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austri
  Methods: serological methods, genetic methods
  
   
  
  Chlamydophila psittaci
  CSDB ID 20491 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: OCH2CH=CH2 or BSA
Trivial name: neoglycoconjugate
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 3163
  Müller-Loennies S, Gronow S, Brade L, Mackenzie R, Kosma P, Brade H "A monoclonal antibody against a carbohydrate epitope in lipopolysaccharide differentiates Chlamydophila psittaci from Chlamydophila pecorum, Chlamydophila pneumoniae, and Chlamydia trachomatis" - Glycobiology 16(3) (2006) 184-196
  

  Lipopolysaccharide (LPS) of Chlamydophila psittaci but not of Chlamydophila pneumoniae or Chlamydia trachomatis contains a tetrasaccharide of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) of the sequence Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo. After immunization with the synthetic neoglycoconjugate antigen Kdo(2→8)[Kdo(2→4)]Kdo(2→4) Kdo-BSA, we obtained the mouse monoclonal antibody (mAb) S69-4 which was able to differentiate C. psittaci from Chlamydophila pecorum, C. pneumoniae, and C. trachomatis in double labeling experiments of infected cell monolayers and by enzyme-linked immunosorbent assay (ELISA). The epitope specificity of mAb S69-4 was determined by binding and inhibition assays using bacteria, LPS, and natural or synthetic Kdo oligosaccharides as free ligands or conjugated to BSA. The mAb bound preferentially Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo(2→4) with a K(d) of 10 microM, as determined by surface plasmon resonance (SPR) for the monovalent interaction using mAb or single chain Fv. Cross-reactivity was observed with Kdo(2→4)Kdo(2→4)Kdo but not with Kdo(2→8)Kdo(2→4)Kdo, Kdo disaccharides in 2→4- or 2→8-linkage, or Kdo monosaccharide. MAb S69-4 was able to detect LPS on thin-layer chromatography (TLC) plates in amounts of <10 ng by immunostaining. Due to the high sensitivity achieved in this assay, the antibody also detected in vitro products of cloned Kdo transferases of Chlamydia. The antibody can therefore be used in medical and veterinarian diagnostics, general microbiology, analytical biochemistry, and studies of chlamydial LPS biosynthesis. Further contribution to the general understanding of carbohydrate-binding antibodies was obtained by a comparison of the primary structure of mAb S69-4 to that of mAb S45-18 of which the crystal structure in complex with its ligand has been elucidated recently (Nguyen et al., 2003, Nat. Struct. Biol., 10, 1019-1025).

  Kdo, diagnostic, neoglycoconjugate, immunofluorescence

  NCBI PubMed ID: 16282606
  Publication DOI: 10.1093/glycob/cwj055
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany, Institute for Biological Sci-ences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austri
  Methods: serological methods, genetic methods
  
   
  
  Chlamydophila psittaci
  CSDB ID 20497 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: BSA
Trivial name: neoglycoconjugate
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_130662,IEDB_135394,IEDB_135813,IEDB_137340,IEDB_140092,IEDB_141807,IEDB_150077,IEDB_150755,IEDB_150756,IEDB_150760,IEDB_150901,IEDB_150908,IEDB_151531,IEDB_151767,IEDB_151768,IEDB_151769,IEDB_156486,IEDB_156487,IEDB_164045

• Article ID: 3163
  Müller-Loennies S, Gronow S, Brade L, Mackenzie R, Kosma P, Brade H "A monoclonal antibody against a carbohydrate epitope in lipopolysaccharide differentiates Chlamydophila psittaci from Chlamydophila pecorum, Chlamydophila pneumoniae, and Chlamydia trachomatis" - Glycobiology 16(3) (2006) 184-196
  

  Lipopolysaccharide (LPS) of Chlamydophila psittaci but not of Chlamydophila pneumoniae or Chlamydia trachomatis contains a tetrasaccharide of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid (Kdo) of the sequence Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo. After immunization with the synthetic neoglycoconjugate antigen Kdo(2→8)[Kdo(2→4)]Kdo(2→4) Kdo-BSA, we obtained the mouse monoclonal antibody (mAb) S69-4 which was able to differentiate C. psittaci from Chlamydophila pecorum, C. pneumoniae, and C. trachomatis in double labeling experiments of infected cell monolayers and by enzyme-linked immunosorbent assay (ELISA). The epitope specificity of mAb S69-4 was determined by binding and inhibition assays using bacteria, LPS, and natural or synthetic Kdo oligosaccharides as free ligands or conjugated to BSA. The mAb bound preferentially Kdo(2→8)[Kdo(2→4)]Kdo(2→4)Kdo(2→4) with a K(d) of 10 microM, as determined by surface plasmon resonance (SPR) for the monovalent interaction using mAb or single chain Fv. Cross-reactivity was observed with Kdo(2→4)Kdo(2→4)Kdo but not with Kdo(2→8)Kdo(2→4)Kdo, Kdo disaccharides in 2→4- or 2→8-linkage, or Kdo monosaccharide. MAb S69-4 was able to detect LPS on thin-layer chromatography (TLC) plates in amounts of <10 ng by immunostaining. Due to the high sensitivity achieved in this assay, the antibody also detected in vitro products of cloned Kdo transferases of Chlamydia. The antibody can therefore be used in medical and veterinarian diagnostics, general microbiology, analytical biochemistry, and studies of chlamydial LPS biosynthesis. Further contribution to the general understanding of carbohydrate-binding antibodies was obtained by a comparison of the primary structure of mAb S69-4 to that of mAb S45-18 of which the crystal structure in complex with its ligand has been elucidated recently (Nguyen et al., 2003, Nat. Struct. Biol., 10, 1019-1025).

  Kdo, diagnostic, neoglycoconjugate, immunofluorescence

  NCBI PubMed ID: 16282606
  Publication DOI: 10.1093/glycob/cwj055
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz Center for Medicine and Biosciences, Borstel, Germany, Institute for Biological Sci-ences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6, Department of Chemistry, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austri
  Methods: serological methods, genetic methods
  
   
  
  Escherichia coli, Chlamydophila psittaci
  CSDB ID 20498 (all data & tools)

Show legend Show as text

Structure type: oligomer
Aglycon: lipid A-BSA
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 3849
  Gerstenbruch S, Brooks CL, Kosma P, Brade L, MacKenzie CR, Evans SV, Brade H, Müller-Loennies S "Analysis of Cross-Reactive and Specific Anti-Carbohydrate Antibodies against Lipopolysaccharide from Chlamydophila psittaci" - Glycobiology 20(4) (2010) 461-472
  

  Chlamydiae contain a rough type lipopolysaccharide (LPS) of 3-deoxy-α-D-manno-oct-2-ulopyranosonic acid residues (Kdo). Two Kdo trisaccharides, 2.8/2.4- and 2.4/2.4-linked, and a branched 2.4[2.8]2.4-linked Kdo tetrasaccharide occur in Chlamydiaceae. While the 2.8/2.4-linked trisaccharide contains a family-specific epitope, the branched Kdo oligosaccharide occurs only in Chlamydophila psittaci and antibodies against it will be useful in human and veterinarian diagnostics. To overcome the generation of cross-reactive antibodies that bind with high affinity to a dominant epitope formed by 2.4/2.4-linked Kdo, we immunized mice with a synthetic 2.4[2.8]-linked branched Kdo trisaccharide and used phage display of scFv to isolate recombinant antibody fragments (NH2240-31 and SAG506-01) that recognize the branched Kdo oligosaccharide with a K(D) of less than 10 nM. Importantly, although these antibodies used germline genes coding for an inherited Kdo recognition site they were able clearly to distinguish between 2.4[2.8]2.4- and 2.4/2.4-linked Kdo. Sequence determination, binding data, and X-ray structural analysis revealed the basis for the improved discrimination between similar Kdo ligands and indicated that the alteration of a stacking interaction from a phenylalanine residue in the center of the combining site to a tyrosine residue facing away from the center favours recognition of branched 2.4[2.8]2.4-linked Kdo residues. Immunofluorescence tests of infected cell monolayers using this antibody show specific staining of C. psittaci elementary bodies that allow it to be distinguished from other pathogenic chlamydiae.

  carbohydrate, Kdo, antibody, Chlamydia, specificity

  NCBI PubMed ID: 20022906
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: sml@fz-borstel.de
  Institutions: Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany
  Methods: X-ray, ELISA, serological methods, genetic methods, surface plasmon resonance (SPR), immunofluorescence analyses, fluorescence microscopy
  
   
  
  Chlamydophila psittaci
  CSDB ID 25652 (all data & tools)

Show legend Show as text

Structure type: oligomer
Trivial name: cross-reactive epitope
Compound class: core oligosaccharide, LPS
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_135394,IEDB_137340,IEDB_140092,IEDB_141807,IEDB_150755,IEDB_150756,IEDB_150760,IEDB_150901,IEDB_150908,IEDB_151531,IEDB_151769,IEDB_176772,IEDB_534865

• Article ID: 4014
  Evans DW, Müller-Loennies S, Brooks CL, Brade L, Kosma P, Brade H, Evans SV "Structural insights into parallel strategies for germline antibody recognition of lipopolysaccharide from Chlamydia" - Glycobiology 21(8) (2011) 1049-1059
  

  The structure of the antigen-binding fragment from the monoclonal antibody S64-4 in complex with a pentasaccharide bisphosphate fragment from chlamydial lipopolysaccharide has been determined by x-ray diffraction to 2.6 A resolution. Like the well-characterized antibody S25-2, S64-4 displays a pocket formed by the residues of germline sequence corresponding to the heavy and light chain V gene segments that binds the terminal Kdo residue of the antigen; however, although S64-4 shares the same heavy chain V gene segment as S25-2, it has a different light chain V gene segment. The new light chain V gene segment codes for a combining site that displays greater affinity, different specificity, and allows a novel antigen conformation that brings a greater number of antigen residues into the combining site than possible in S25-2. Further, while antibodies in the S25-2 family use complementarity determining region (CDR) H3 to discriminate among antigens, S64-4 achieves its specificity via the new light chain V gene segment and resulting change in antigen conformation. These structures reveal an intriguing parallel strategy where two different combinations of germline-coded V gene segments can act as starting points for the generation of germline antibodies against chlamydial antigens and show how anti-carbohydrate antibodies can exploit the conformational flexibility of this class of antigens to achieve high affinity and specificity independently of CDR H3.

  Chlamydia, crystal structure, carbohydrate antigen, antibody-antigen recognition, chlamydial LPS, germline antibody

  NCBI PubMed ID: 21543444
  Publication DOI: 10.1093/glycob/cwr041
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: hebra@fz-borstel.de; svenvans@uvic.ca
  Institutions: Department of Biochemistry and Microbiology, University of Victoria, PO Box 3055 STN CSC, Victoria, BC, Canada V8P 3P6, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 22, Borstel D-23845, Germany, Department of Chemistry, University of Natural Resources and Life Sciences, Vienna A-1190, Austria
  Methods: X-ray, ELISA, serological methods
  
   
  
  Chlamydia psittaci
  CSDB ID 26190 (all data & tools)
• Article ID: 4677
  Haji-Ghassemi O, Müller-Loennies S, Saldova R, Muniyappa M, Brade L, Rudd PM, Harvey DJ, Kosma P, Brade H, Evans SV "Groove-type recognition of Chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan" - Journal of Biological Chemistry 289(24) (2014) 16644-16661
  

  The structure of the antigen-binding fragment of mAb S25-26 determined to 1.95 A resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)-Kdo(2→4)Kdo (Kdo=3-deoxy-α-D-manno-oct-2-ulopyranosonic acid) displays a germline-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs, despite being raised against the identical immunogen. Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germline residues to recognize the entire trisaccharide antigen and so confer strict specificity. Interest in S25-23 was sparked by its rare high muM affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. The structures of three unliganded forms of S25-26 have a labile complementary determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. The elucidation of the three dimensional structure of an αGal containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as they have been implicated in allergic response in patients receiving therapeutic antibodies.

  antibodies, epitopes, MAb, Chlamydiaceae, N-linked glycan, lipopolysaccharide antigen

  NCBI PubMed ID: 24682362
  Publication DOI: 10.1074/jbc.M113.528224
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: sml@fz-borstel.de; svevans@uvic.ca
  Institutions: From the Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8P 3P6, Canada, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 22, Borstel D-23845, Germany
  Methods: X-ray, ELISA, ESI-MS, genetic methods, HPLC, crystallization, ITC
  
   
  
  Chlamydia psittaci 6BC
  CSDB ID 30245 (all data & tools)

Show legend Show as text

Structure type: oligomer
Trivial name: cross-reactive epitope
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 4677
  Haji-Ghassemi O, Müller-Loennies S, Saldova R, Muniyappa M, Brade L, Rudd PM, Harvey DJ, Kosma P, Brade H, Evans SV "Groove-type recognition of Chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan" - Journal of Biological Chemistry 289(24) (2014) 16644-16661
  

  The structure of the antigen-binding fragment of mAb S25-26 determined to 1.95 A resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)-Kdo(2→4)Kdo (Kdo=3-deoxy-α-D-manno-oct-2-ulopyranosonic acid) displays a germline-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs, despite being raised against the identical immunogen. Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germline residues to recognize the entire trisaccharide antigen and so confer strict specificity. Interest in S25-23 was sparked by its rare high muM affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. The structures of three unliganded forms of S25-26 have a labile complementary determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. The elucidation of the three dimensional structure of an αGal containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as they have been implicated in allergic response in patients receiving therapeutic antibodies.

  antibodies, epitopes, MAb, Chlamydiaceae, N-linked glycan, lipopolysaccharide antigen

  NCBI PubMed ID: 24682362
  Publication DOI: 10.1074/jbc.M113.528224
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: sml@fz-borstel.de; svevans@uvic.ca
  Institutions: From the Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8P 3P6, Canada, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 22, Borstel D-23845, Germany
  Methods: X-ray, ELISA, ESI-MS, genetic methods, HPLC, crystallization, ITC
  
   
  
  Chlamydia psittaci
  CSDB ID 30244 (all data & tools)

Show legend Show as text

Structure type: fragment of a bigger structure
Trivial name: Chlamydia epitope
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 4861
  Pokorny B, Kosma P "Synthesis of Chlamydia lipopolysaccharide haptens through the use of a-specific 3-iodo-Kdo fluoride glycosyl donors" - Chemistry 21(1) (2015) 305-313
  

  A scalable approach towards high-yielding and (stereo)selective glycosyl donors of the 2-ulosonic acid Kdo (3-deoxy-D-manno-oct-2-ulosonic acid) is a fundamental requirement for the development of vaccines against Gram-negative bacteria. Herein, we disclose a short synthetic route to 3-iodo Kdo fluoride donors from Kdo glycal esters that enable efficient alpha-specific glycosylations and significantly suppress the elimination side reaction. The potency of these donors is demonstrated in a straightforward, six-step synthesis of a branched Chlamydia-related Kdo-trisaccharide ligand without the need for protecting groups at the Kdo glycosyl acceptor. The approach was further extended to include sequential iteration of the basic concept to produce the linear Chlamydia-specific α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo trisaccharide in a good overall yield.

  carbohydrates, 3-deoxy-D-manno-oct-2-ulosonic acid, Kdo, Oligosaccharides, Chlamydia, glycosylation, immunochemistry, diastereoselectivity

  NCBI PubMed ID: 25354167
  Publication DOI: 10.1002/chem.201405424
  Publisher: Vch Verlagsgesellschaft
  Correspondence: paul.kosma@boku.ac.at
  Institutions: Department of Chemistry, University of Natural Resources and Life Sciences-Vienna, Muthgasse 18, 1190 Vienna (Austria)
  Methods: 13C NMR, 1H NMR, TLC, chemical synthesis, chemical methods, ESI-TOF-MS
  
   
  
  Chlamydia psittaci
  CSDB ID 11201 (all data & tools)

Show legend Show as text

Structure type: fragment of a bigger structure
Aglycon: (2->6)lipid A
Trivial name: Kdo-antigen of Chlamydiaceae
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_150756,IEDB_150901,IEDB_151769

• Article ID: 4972
  Kosma P "Progress in Kdo-glycoside chemistry" - Tetrahedron Letters 57(20) (2016) 2133-2142
  

  Glycosylation chemistry of 3-deoxy-D-manno-oct-2-ulosonic acid units has been considerably developed within the last decade. This review covers major achievements with respect to improved yields and anomeric selectivity as well as suppression of the elimination side reaction via selection of dedicated protecting groups and appropriate activation of the anomeric center.

  synthesis, oligosaccharide, 3-deoxy-D-manno-oct-2-ulosonic acid, Kdo, glycosyl donor, Glycal

  Publication DOI: 10.1016/j.tetlet.2016.04.005
  Journal NLM ID: 2984819R
  Publisher: Elsevier
  Correspondence: paul.kosma@boku.ac.at
  Institutions: University of Natural Resources and Life Sciences-Vienna, Department of Chemistry, Muthgasse 18, A-1190 Vienna, Austria
  
   
  
  (Chlamydiaceae)
  CSDB ID 11851 (all data & tools)