Show legend Show as text

Structure type: monomer
Contained glycoepitopes: IEDB_130650,IEDB_141494,IEDB_167475

• Article ID: 133
  Tzeng YL, Datta A, Strole CA, Birck MR, Taylor WP, Carlson RW, Woodard RW, Stephens DS "KpsF is the arabinose 5-phosphate isomerase required for 3-deoxy-D-manno-octulosonic acid (Kdo) biosynthesis and for both lipooligosaccharide assembly and capsular polysaccharide expression in Neisseria meningitidis" - Journal of Biological Chemistry 277(27) (2002) 24103-24113
  

  We have identified and defined the function of kpsF of Neisseria meningitidis, and the homologues of kpsF in encapsulated K1 and K5 Escherichia coli. KpsF was shown to be the arabinose 5-phosphate isomerase, an enzyme not previously identified in prokaryotes that mediates the interconversion of ribulose 5-phosphate and arabinose 5- phosphate. KpsF is required for 3-deoxy-D-manno-octulosonic acid (Kdo) biosynthesis in N. meningitidis. Mutation of kpsF or the gene encoding the CMP-Kdo synthetase (kpsU/kdsB) in N. meningitidis resulted in expression of a lipooligossaccharide (LOS) structure that contained only lipid A and reduced capsule expression in the five invasive disease associated meningococcal serogroups (A, B, C, Y, and W-135). The step linking meningococcal capsule and LOS biosynthesis was shown to be Kdo production as the expression of capsule was wild type in a Kdo transferase (kdtA) mutant. Thus, in addition to lipooligosaccharide assembly, Kdo is required for meningococcal capsular polysaccharide expression. Further, N. meningitidis, unlike enteric gram-negative bacteria, can survive and synthesize only unglycosylated lipid A

  biosynthesis, structure, Lipooligosaccharide, Neisseria meningitidis, disease, expression, gene, invasive, LOS, meningococcal, Neisseria, capsular, polysaccharide, capsular polysaccharide, Escherichia, Escherichia coli, acid, Kdo, transferase, lipid, lipid A, phosphate, type, wild type, infectious disease, bacteria, mutant, serogroup, assembly, production, reduced, medicine, Gram-negative bacteria, 3-deoxy-D-manno-octulosonic acid, arabinose, capsule, defined, enzyme, function, gram negative bacteria, Gram-negative, Infectious, interconversion, mutation, synthetase

  NCBI PubMed ID: 11956197
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Institutions: Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine, Atlanta, GA 30303
  Methods: biochemical methods
  
   
  
  Escherichia coli
  CSDB ID 5409 (all data & tools)

Show legend Show as text

Structure type: monomer
Contained glycoepitopes: IEDB_141494,IEDB_167475,IEDB_838988

• Article ID: 3239
  Shoenhofen IC, McNally DJ, Brisson JR, Logan SM "Elucidation of the CMP-pseudaminic acid pathway in Helicobacter pylori: synthesis from UDP-N-acetylglucosamine by a single enzymatic reaction" - Glycobiology 16(9) (2006) 8C-14C
  

  Flagellin glycosylation is a necessary modification allowing flagellar assembly, bacterial motility, colonization, and hence virulence for the gastrointestinal pathogen Helicobacter pylori [Josenhans, C., Vossebein, L., Friedrich, S., and Suerbaum, S. (2002) FEMS Microbiol. Lett., 210, 165-172; Schirm, M., Schoenhofen, I.C., Logan, S.M., Waldron, K.C., and Thibault, P. (2005) Anal. Chem., 77, 7774-7782]. A causative agent of gastric and duodenal ulcers, H. pylori, heavily modifies its flagellin with the sialic acid-like sugar 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-α-L-manno-nonulosonic acid (pseudaminic acid). Because this sugar is unique to bacteria, its biosynthetic pathway offers potential as a novel therapeutic target. We have identified six H. pylori enzymes, which reconstitute the complete biosynthesis of pseudaminic acid, and its nucleotide-activated form CMP-pseudaminic acid, from UDP-N-acetylglucosamine (UDP-GlcNAc). The pathway intermediates and final product were identified from monitoring sequential reactions with nuclear magnetic resonance (NMR) spectroscopy, thereby confirming the function of each biosynthetic enzyme. Remarkably, the conversion of UDP-GlcNAc to CMP-pseudaminic acid was achieved in a single reaction combining six enzymes. This represents the first complete in vitro enzymatic synthesis of a sialic acid-like sugar and sets the groundwork for future small molecule inhibitor screening and design. Moreover, this study provides a strategy for efficient large-scale synthesis of novel medically relevant bacterial sugars that has not been attainable by chemical methods alone.

  sialic acid, Helicobacter pylori, enzymatic synthesis, biosynthetic pathway elucidation, CMP-pseudaminic acid, flagellin glycosylation

  NCBI PubMed ID: 16751642
  Publication DOI: 10.1093/glycob/cwl010
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: susan.logan@nrc-cnrc.gc.ca
  Institutions: Institute for Biological Sciences, National Research Council, Ottawa,Ontario, Canada K1A OR6
  Methods: NMR, biochemical methods
  
   
  
  Helicobacter pylori
  CSDB ID 20349 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: sialoside
Contained glycoepitopes: IEDB_136794,IEDB_141494,IEDB_146100,IEDB_149174,IEDB_167475,SB_170,SB_171,SB_172,SB_84

• Article ID: 3316
  Yu H, Chen X "Carbohydrate post-glycosylational modifications" - Organic and Biomolecular Chemistry 5(6) (2007) 865-872
  

  Carbohydrate modification is a common phenomenon in nature. Many carbohydrate modifications such as some epimerization, O-acetylation, O-sulfation, O-methylation, N-deacetylation, and N-sulfation, take place after the formation of oligosaccharide or polysaccharide backbones. These modifications can be categorized as carbohydrate post-glycosylational modifications (PGMs). Carbohydrate PGMs further extend the complexity of the structures and the synthesis of carbohydrates and glycoconjugates. They also increase the capacity of the biological regulation that is achieved by finely tuning the structures of carbohydrates. Developing efficient methods to obtain structurally defined naturally occurring oligosaccharides, polysaccharides, and glycoconjugates with carbohydrate PGMs is essential for understanding the biological significance of carbohydrate PGMs. Combined with high-throughput screening methods, synthetic carbohydrates with PGMs are invaluable probes in structure-activity relationship studies. We illustrate here several classes of carbohydrates with PGMs and their applications. Recent progress in chemical, enzymatic, and chemoenzymatic syntheses of these carbohydrates and their derivatives are also presented

  synthesis, polysaccharides, regulation, glycoconjugates, enzymatic, modification, structure-activity relationship, methods, epimerization

  Publication DOI: 10.1039/b700034k
  Journal NLM ID: 101154995
  Publisher: The Royal Society of Chemistry
  Correspondence: chen@chem.ucdavis.edu
  Institutions: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA
  
   
  
  Campylobacter jejuni
  CSDB ID 21409 (all data & tools)

Show legend Show as text

Structure type: monomer
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3325
  Vionnet J, Vann WF "Successive glycosyltransfer of sialic acid by Escherichia coli K92 polysialyltransferase in elongation of oligosialic acceptors" - Glycobiology 17(7) (2007) 735-743
  

  Escherichia coli K92 produces a capsular polysialic acid with alternating α2,8 α2,9 NeuNAc linkages. This polysaccharide is cross reactive with the neuroinvasive pathogen Neisseria meningitidis Group C. The K92 polysialyltransferase catalyzes the synthesis of the polysialic acid with alternating linkages by the transfer of NeuNAc from CMP-NeuNAc to the non-reducing end of the growing polymer. We used a fluorescent based HPLC assay to characterize the process of chain extension. The polysialyltransferase elongates the acceptor GT3-FCHASE in a biphasic fashion. The initial phase polymers are characterized by accumulation of product containing 1 to 8 additional sialic acid residues. This phase is followed by a very rapid formation of high molecular weight polymer as the accumulated oligosaccharides containing 8-10 sialic acids are consumed. The high molecular weight polymer contains 90-100 sialic acids and is sensitive to degradation by periodate and K1-5 endoneuraminidase suggesting that the polymer contains the alternating structure. The polymerization reaction does not appear to be strictly processive, since oligosaccharides of each intermediate size were detected before accumulation of high molecular weight polymer. Synthesis can be blocked by CMP-9-azido-NeuNAc. These results suggest that the K92 polysialyltransferase forms both α2,8 and α2,9 linkages in a successive and non-processive fashion

  capsular polysaccharides, sialic acid, polysialyltransferase, chain extension, processivity

  NCBI PubMed ID: 17384120
  Publication DOI: 10.1093/glycob/cwm032
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: wvann@helix.nih.gov
  Institutions: Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, FDA, Bethesda, MD, USA
  Methods: biochemical methods, HPLC
  
   
  
  Escherichia coli K92
  CSDB ID 21425 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-α-L-manno-non-2-ulosonic (CMP-Pse5Ac7AC), CMP-pseudaminic acid
Contained glycoepitopes: IEDB_141494,IEDB_167475,IEDB_838988

• Article ID: 3370
  McNally DJ, Aubry AJ, Hui JP, Khieu NH, Whitfield D, Ewing CP, Guerry P, Brisson JR, Logan SM, Soo EC "Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans" - Journal of Biological Chemistry 282(19) (2007) 14463-14475
  

  Glycosylation of Campylobacter flagellin is required for the biogenesis of a functional flagella filament. Recently, we used a targeted metabolomics approach using mass spectrometry and NMR to identify changes in the metabolic profile of wild type and mutants in the flagellar glycosylation locus, characterize novel metabolites, and assign function to genes to define the pseudaminic acid biosynthetic pathway in Campylobacter jejuni 81-176 (McNally, D. J., Hui, J. P., Aubry, A. J., Mui, K. K., Guerry, P., Brisson, J. R., Logan, S. M., and Soo, E. C. (2006) J. Biol. Chem. 281, 18489-18498). In this study, we use a similar approach to further define the glycome and metabolomic complement of nucleotide-activated sugars in Campylobacter coli VC167. Herein we demonstrate that, in addition to CMP-pseudaminic acid, C. coli VC167 also produces two structurally distinct nucleotide-activated nonulosonate sugars that were observed as negative ions at m/z 637 and m/z 651 (CMP-315 and CMP-329). Hydrophilic interaction liquid chromatography-mass spectrometry yielded suitable amounts of the pure sugar nucleotides for NMR spectroscopy using a cold probe. Structural analysis in conjunction with molecular modeling identified the sugar moieties as acetamidino and N-methylacetimidoyl derivatives of legionaminic acid (Leg5Am7Ac and Leg5AmNMe7Ac). Targeted metabolomic analyses of isogenic mutants established a role for the ptmA-F genes and defined two new ptm genes in this locus as legionaminic acid biosynthetic enzymes. This is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein

  metabolism, glycan, legionaminic acid, modeling, Flagellin, Campylobacter coli

  NCBI PubMed ID: 17371878
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: susan.logan@nrc-cnrc.gc.ca; evelyn.soo@nrc-cnrc.gc.ca
  Institutions: National Research Council, Institute for Biological Sciences, Ottawa, Ontario, Canada
  Methods: 13C NMR, 1H NMR, NMR-2D, MD simulations, NMR-1D, genetic methods, biochemical methods, biosynthetic methods, CE-ESI-MS, LC-MS
  
   
  
  Campylobacter coli VC167
  CSDB ID 21505 (all data & tools)
• Article ID: 3766
  Rangarajan ES, Proteau A, Cui Q, Logan SM, Potetinova Z, Whitfield D, Purisima EO, Cygler M, Matte A, Sulea T, Schoenhofen IC "Structural and Functional Analysis of Campylobacter jejuni PseG: A UDP-sugar hydrolase from the pseudaminic acid biosynthetic pathway" - Journal of Biological Chemistry 284(31) (2009) 20989-21000
  

  Flagella of the bacteria Helicobacter pylori and Campylobacter jejuni are important virulence determinants, whose proper assembly and function are dependent upon glycosylation at multiple positions by sialic acid-like sugars, such as 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno-nonulosonic acid (pseudaminic acid (Pse)). The fourth enzymatic step in the pseudaminic acid pathway, the hydrolysis of UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose to generate 2,4-diacetamido-2,4,6-trideoxy-l-altropyranose, is performed by the nucleotide sugar hydrolase PseG. To better understand the molecular basis of the PseG catalytic reaction, we have determined the crystal structures of C. jejuni PseG in apo-form and as a complex with its UDP product at 1.8 and 1.85 A resolution, respectively. In addition, molecular modeling was utilized to provide insight into the structure of the PseG-substrate complex. This modeling identifies a His(17)-coordinated water molecule as the putative nucleophile and suggests the UDP-sugar substrate adopts a twist-boat conformation upon binding to PseG, enhancing the exposure of the anomeric bond cleaved and favoring inversion at C-1. Furthermore, based on these structures a series of amino acid substitution derivatives were constructed, altering residues within the active site, and each was kinetically characterized to examine its contribution to PseG catalysis. In conjunction with structural comparisons, the almost complete inactivation of the PseG H17F and H17L derivatives suggests that His(17) functions as an active site base, thereby activating the nucleophilic water molecule for attack of the anomeric C-O bond of the UDP-sugar. As the PseG structure reveals similarity to those of glycosyltransferase family-28 members, in particular that of Escherichia coli MurG, these findings may also be of relevance for the mechanistic understanding of this important enzyme family.

  Campylobacter jejuni, pseudaminic acid, crystal structure, Helicobacter pylori, glycosyltransferase, modeling, hydrolase, flagella

  NCBI PubMed ID: 19483088
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: traian.sulea@nrc-cnrc.gc.ca; ian.schoenhofen@nrc-cnrc.gc.ca
  Institutions: From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1V6
  Methods: X-ray, SDS-PAGE, MD simulations, genetic methods, molecular modeling, CD, crystallization
  
   
  
  Campylobacter jejuni
  CSDB ID 24194 (all data & tools)
• Article ID: 4769
  Li Z, Hwang S, Ericson J, Bowler K, Bar-Peled M "Pen and Pal Are Nucleotide-Sugar Dehydratases That Convert UDP-GlcNAc to UDP-6-Deoxy-D-GlcNAc-5,6-ene and Then to UDP-4-Keto-6-deoxy-L-AltNAc for CMP-Pseudaminic Acid Synthesis in Bacillus thuringiensis" - Journal of Biological Chemistry 290(2) (2015) 691-704
  

  CMP-pseudaminic acid is a precursor required for the O-glycosylation of flagellin in some pathogenic Gram-negative bacteria, a process known to be critical in bacterial motility and infection. However, little is known about flagellin glycosylation in Gram-positive bacteria. Here, we identified and functionally characterized an operon, named Bti_pse, in Bacillus thuringiensis israelensis ATCC 35646, which encodes seven different enzymes that together convert UDP-GlcNAc to CMP-pseudaminic acid. In contrast, Gram-negative bacteria complete this reaction with six enzymes. The first enzyme, which we named Pen, converts UDP-D-GlcNAc to an uncommon UDP-sugar, UDP-6-deoxy-D-GlcNAc-5,6-ene. Pen contains strongly bound NADP+ and has distinct UDP-GlcNAc 4-oxidase, 5,6-dehydratase, and 4-reductase activities. The second enzyme, which we named Pal, converts UDP-6-deoxy-D-GlcNAc-5,6-ene to UDP-4-keto-6-deoxy-L-AltNAc. Pal is NAD+-dependent and has distinct UDP-6-deoxy-D-GlcNAc-5,6-ene 4-oxidase, 5,6-reductase, and 5-epimerase activities. We also show here using NMR spectroscopy and mass spectrometry that in B. thuringiensis, the enzymatic product of Pen and Pal, UDP-4-keto-6-deoxy-L-AltNAc, is converted to CMP-pseudaminic acid by the sequential activities of a C4?-transaminase (Pam), a 4-N-acetyltransferase (Pdi), a UDP-hydrolase (Phy), an enzyme (Ppa) that adds phosphoenolpyruvate to form pseudaminic acid, and finally a cytidylyltransferase that condenses CTP to generate CMP-pseudaminic acid. Knowledge of the distinct dehydratase-like enzymes Pen and Pal and their role in CMP-pseudaminic acid biosynthesis in Gram-positive bacteria provides a foundation to investigate the role of pseudaminic acid and flagellin glycosylation in Bacillus and their involvement in bacterial motility and pathogenicity.

  pseudaminic acid, Bacillus, aminotransferase, CMP-pseudaminic acid, flagellin glycosylation, dehydratase, enzyme catalysis, bacterial metabolism, Bacillus thuringiensis, carbohydrate biosynthesis, glycobiology, UDP-GlcNAc

  NCBI PubMed ID: 25414257
  Publication DOI: 10.1074/jbc.M114.612747
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: peled@ccrc.uga.edu
  Institutions: Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA, From the Department of Plant Biology and Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA
  Methods: gel filtration, 13C NMR, 1H NMR, NMR-2D, PCR, SDS-PAGE, DNA techniques, kinetics assays, NMR-1D, genetic methods, biochemical methods, HPLC, UV, enzymatic analysis, LC-ESI-MS/MS, HILIC-HPLC-UV, HILIC-ESI-MS
  
   
  
  Bacillus thuringiensis sv. israelensis ATCC 35646
  CSDB ID 30629 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto-non-2-ulosonic (CMP-Leg5Ac7AC), CMP-5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-β-D-galacto-non-2-ulosonic acid, CMP-legionaminic acid, CMP-N,N'-diacetyllegionaminic acid, CMP-Leg5Ac7Ac
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3370
  McNally DJ, Aubry AJ, Hui JP, Khieu NH, Whitfield D, Ewing CP, Guerry P, Brisson JR, Logan SM, Soo EC "Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans" - Journal of Biological Chemistry 282(19) (2007) 14463-14475
  

  Glycosylation of Campylobacter flagellin is required for the biogenesis of a functional flagella filament. Recently, we used a targeted metabolomics approach using mass spectrometry and NMR to identify changes in the metabolic profile of wild type and mutants in the flagellar glycosylation locus, characterize novel metabolites, and assign function to genes to define the pseudaminic acid biosynthetic pathway in Campylobacter jejuni 81-176 (McNally, D. J., Hui, J. P., Aubry, A. J., Mui, K. K., Guerry, P., Brisson, J. R., Logan, S. M., and Soo, E. C. (2006) J. Biol. Chem. 281, 18489-18498). In this study, we use a similar approach to further define the glycome and metabolomic complement of nucleotide-activated sugars in Campylobacter coli VC167. Herein we demonstrate that, in addition to CMP-pseudaminic acid, C. coli VC167 also produces two structurally distinct nucleotide-activated nonulosonate sugars that were observed as negative ions at m/z 637 and m/z 651 (CMP-315 and CMP-329). Hydrophilic interaction liquid chromatography-mass spectrometry yielded suitable amounts of the pure sugar nucleotides for NMR spectroscopy using a cold probe. Structural analysis in conjunction with molecular modeling identified the sugar moieties as acetamidino and N-methylacetimidoyl derivatives of legionaminic acid (Leg5Am7Ac and Leg5AmNMe7Ac). Targeted metabolomic analyses of isogenic mutants established a role for the ptmA-F genes and defined two new ptm genes in this locus as legionaminic acid biosynthetic enzymes. This is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein

  metabolism, glycan, legionaminic acid, modeling, Flagellin, Campylobacter coli

  NCBI PubMed ID: 17371878
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: susan.logan@nrc-cnrc.gc.ca; evelyn.soo@nrc-cnrc.gc.ca
  Institutions: National Research Council, Institute for Biological Sciences, Ottawa, Ontario, Canada
  Methods: 13C NMR, 1H NMR, NMR-2D, MD simulations, NMR-1D, genetic methods, biochemical methods, biosynthetic methods, CE-ESI-MS, LC-MS
  
   
  
  Campylobacter coli VC167
  CSDB ID 21874 (all data & tools)
• Article ID: 3474
  Glaze PA, Watson DC, Young NM, Tanner ME "Biosynthesis of CMP-N,N'-diacetyllegionaminic acid from UDP-N,N'-diacetylbacillosamine in Legionella pneumophila" - Biochemistry 47(10) (2008) 3272-3282
  

  Legionaminic acid is a nine-carbon α-keto acid that is similar in structure to other members of the sialic acid family that includes neuraminic acid and pseudaminic acid. It is found as a component of the lipopolysaccharide in several bacterial species and is perhaps best known for its presence in the O-antigen of the causative agent of Legionnaires' disease, Legionella pneumophila. In this work, the enzymes responsible for the biosynthesis and activation of N, N'-diacetyllegionaminic acid are identified for the first time. A cluster of three L. pneumophila genes bearing homology to known sialic acid biosynthetic genes (neuA,B,C) were cloned and overexpressed in Escherichia coli. The NeuC homologue was found to be a hydrolyzing UDP- N, N'-diacetylbacillosamine 2-epimerase that converts UDP- N, N'-diacetylbacillosamine into 2,4-diacetamido-2,4,6-trideoxymannose and UDP. Stereochemical and isotopic labeling studies showed that the enzyme utilizes a mechanism involving an initial anti elimination of UDP to form a glycal intermediate and a subsequent syn addition of water to generate product. This is similar to the hydrolyzing UDP- N-acetylglucosamine 2-epimerase (NeuC) of sialic acid biosynthesis, but the L. pneumophila enzyme would not accept UDP-GlcNAc as an alternate substrate. The NeuB homologue was found to be a N, N'-diacetyllegionaminic acid synthase that condenses 2,4-diacetamido-2,4,6-trideoxymannose with phosphoenolpyruvate (PEP), although the in vitro activity of the recombinant enzyme (isolated as a MalE fusion protein) was very low. The synthase activity was dependent on the presence of a divalent metal ion, and the reaction proceeded via a C-O bond cleavage process, similar to the reactions catalyzed by the sialic acid and pseudaminic acid synthases. Finally, the NeuA homologue was shown to possess the CMP- N, N'-diacetyllegionaminic acid synthetase activity that generates the activated form of legionaminic acid used in lipopolysaccharide biosynthesis. Together, the three enzymes constitute a pathway that converts a UDP-linked bacillosamine derivative into a CMP-linked legionaminic acid derivative

  O-antigen, lipopolysaccharide biosynthesis, pseudaminic acid, neuraminic acid, Legionella pneumophila, legionaminic acid, Legionella, bacillosamine

  NCBI PubMed ID: 18275154
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Correspondence: mtanner@chem.ubc.ca
  Institutions: Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1
  Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, 31P NMR, genetic methods, biochemical methods
  
   
  
  Legionella pneumophila
  CSDB ID 22965 (all data & tools)
• Article ID: 3774
  Schoenhofen IC, Vinogradov E, Whitfield DM, Brisson JR, Logan SM "The CMP-legionaminic acid pathway in Campylobacter: biosynthesis involving novel GDP-linked precursors" - Glycobiology 19(7) (2009) 715-725
  

  The sialic acid-like sugar 5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto-nonulosonic acid, or legionaminic acid, is found as a virulence-associated cell-surface glycoconjugate in the Gram-negative bacteria Legionella pneumophila and Campylobacter coli. L. pneumophila serogroup 1 strains, causative agents of Legionnaire's disease, contain an α2,4-linked homopolymer of legionaminic acid within their lipopolysaccharide O-chains, whereas the gastrointestinal pathogen C. coli modifies its flagellin with this monosaccharide via O-linkage. In this work, we have purified and biochemically characterized eleven candidate biosynthetic enzymes from C. jejuni, thereby fully reconstituting the biosynthesis of legionaminic acid and its CMP-activated form, starting from fructose-6-P. This pathway involves unique GDP-linked intermediates, likely providing a cellular mechanism for differentiating between this and similar UDP-linked pathways, such as UDP-2,4-diacetamido-bacillosamine biosynthesis involved in N-linked protein glycosylation. Importantly, these findings provide a facile method for efficient large-scale synthesis of legionaminic acid, and since legionaminic acid and sialic acid share the same D-glycero-D-galacto absolute configuration, this sugar may now be evaluated for its potential as a sialic acid mimic.

  Campylobacter jejuni, sialic acid, legionaminic acid, flagellin glycosylation, neuraminic aid

  NCBI PubMed ID: 19282391
  Publication DOI: 10.1093/glycob/cwp039
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: ian.schoenhofen@nrc-cnrc-gc.ca
  Institutions: Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada K1A 0R6
  Methods: 13C NMR, 1H NMR, SDS-PAGE, genetic methods, biochemical methods, CE, CE-MS
  
   
  
  Campylobacter jejuni Cj1332
  CSDB ID 24216 (all data & tools)
• Article ID: 4133
  Watson DC, Leclerc S, Wakarchuk WW, Young NM "Enzymatic synthesis and properties of glycoconjugates with legionaminic acid as a replacement for neuraminic acid" - Glycobiology 21(1) (2011) 99-108
  

  In addition to sialic acid, bacteria produce several other nonulosonic acids, including legionaminic acid (Leg). This has exactly the same stereochemistry as sialic acid, with the added features of 9-deoxy and 7-amino groups. In order to explore the biological effects of replacing sialic acid residues (Neu5Ac) in glycoconjugates with Leg in its diacetylated form, diacetyllegionaminic acid (Leg5Ac7Ac), we tested CMP-Leg5Ac7Ac as a donor substrate with a selection of bacterial and mammalian sialyltransferases. The CMP-Leg5Ac7Ac was synthesized in vitro by means of cloned enzymes from the bacillosamine portion of the Campylobacter jejuni N-glycan pathway and from the Leg pathway of Legionella pneumophila. Using fluorescent derivatives of lactose, Galβ1,4GlcNAcβ and T-antigen (Galβ1,3GalNAcα) as acceptors, we tested eight different sialyltransferases and found that the Pasteurella multocida PM0188h and porcine ST3Gal1 sialyltransferases were significantly active with CMP-Leg5Ac7Ac, showing approximately 60% activity when compared with CMP-Neu5Ac. The Photobacterium α2,6 sialyltransferase was weakly active, with approximately 6% relative activity. The Leg5Ac7Ac-α-2,3-lactose product was then tested as a substrate with six sialidases of viral, bacterial and mammalian origin. All showed much lower activities than with the corresponding sialic acid substrate, with the influenza virus N1 being the most active and human NEU2 being the least active. These results show the feasibility of producing glycoconjugates with Leg5Ac7Ac residues as the terminal sugars, which should display novel biological properties.

  sialyltransferase, sialic acid, legionaminic acid, glycoconjugate, neuraminidase

  NCBI PubMed ID: 20978010
  Publication DOI: 10.1093/glycob/cwq135
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: martin.young@nrc-cnrc.gc.ca
  Institutions: Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, Canada
  Methods: DNA cloning, genetic methods, biochemical methods, CE, CE-MS
  
   
  
  Campylobacter jejuni, Legionella pneumophila
  CSDB ID 27009 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-D-glycero-β-D-galacto-non-2-ulosonic (CMP-Leg5Am7Ac)
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3370
  McNally DJ, Aubry AJ, Hui JP, Khieu NH, Whitfield D, Ewing CP, Guerry P, Brisson JR, Logan SM, Soo EC "Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans" - Journal of Biological Chemistry 282(19) (2007) 14463-14475
  

  Glycosylation of Campylobacter flagellin is required for the biogenesis of a functional flagella filament. Recently, we used a targeted metabolomics approach using mass spectrometry and NMR to identify changes in the metabolic profile of wild type and mutants in the flagellar glycosylation locus, characterize novel metabolites, and assign function to genes to define the pseudaminic acid biosynthetic pathway in Campylobacter jejuni 81-176 (McNally, D. J., Hui, J. P., Aubry, A. J., Mui, K. K., Guerry, P., Brisson, J. R., Logan, S. M., and Soo, E. C. (2006) J. Biol. Chem. 281, 18489-18498). In this study, we use a similar approach to further define the glycome and metabolomic complement of nucleotide-activated sugars in Campylobacter coli VC167. Herein we demonstrate that, in addition to CMP-pseudaminic acid, C. coli VC167 also produces two structurally distinct nucleotide-activated nonulosonate sugars that were observed as negative ions at m/z 637 and m/z 651 (CMP-315 and CMP-329). Hydrophilic interaction liquid chromatography-mass spectrometry yielded suitable amounts of the pure sugar nucleotides for NMR spectroscopy using a cold probe. Structural analysis in conjunction with molecular modeling identified the sugar moieties as acetamidino and N-methylacetimidoyl derivatives of legionaminic acid (Leg5Am7Ac and Leg5AmNMe7Ac). Targeted metabolomic analyses of isogenic mutants established a role for the ptmA-F genes and defined two new ptm genes in this locus as legionaminic acid biosynthetic enzymes. This is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein

  metabolism, glycan, legionaminic acid, modeling, Flagellin, Campylobacter coli

  NCBI PubMed ID: 17371878
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: susan.logan@nrc-cnrc.gc.ca; evelyn.soo@nrc-cnrc.gc.ca
  Institutions: National Research Council, Institute for Biological Sciences, Ottawa, Ontario, Canada
  Methods: 13C NMR, 1H NMR, NMR-2D, MD simulations, NMR-1D, genetic methods, biochemical methods, biosynthetic methods, CE-ESI-MS, LC-MS
  
   
  
  Campylobacter coli VC167
  CSDB ID 21875 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-5-E/Z-N-(N-methylacetimidoyl)-7-acetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto-nonulosonic acid, CMP-Leg5AmNMe7Ac
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3370
  McNally DJ, Aubry AJ, Hui JP, Khieu NH, Whitfield D, Ewing CP, Guerry P, Brisson JR, Logan SM, Soo EC "Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans" - Journal of Biological Chemistry 282(19) (2007) 14463-14475
  

  Glycosylation of Campylobacter flagellin is required for the biogenesis of a functional flagella filament. Recently, we used a targeted metabolomics approach using mass spectrometry and NMR to identify changes in the metabolic profile of wild type and mutants in the flagellar glycosylation locus, characterize novel metabolites, and assign function to genes to define the pseudaminic acid biosynthetic pathway in Campylobacter jejuni 81-176 (McNally, D. J., Hui, J. P., Aubry, A. J., Mui, K. K., Guerry, P., Brisson, J. R., Logan, S. M., and Soo, E. C. (2006) J. Biol. Chem. 281, 18489-18498). In this study, we use a similar approach to further define the glycome and metabolomic complement of nucleotide-activated sugars in Campylobacter coli VC167. Herein we demonstrate that, in addition to CMP-pseudaminic acid, C. coli VC167 also produces two structurally distinct nucleotide-activated nonulosonate sugars that were observed as negative ions at m/z 637 and m/z 651 (CMP-315 and CMP-329). Hydrophilic interaction liquid chromatography-mass spectrometry yielded suitable amounts of the pure sugar nucleotides for NMR spectroscopy using a cold probe. Structural analysis in conjunction with molecular modeling identified the sugar moieties as acetamidino and N-methylacetimidoyl derivatives of legionaminic acid (Leg5Am7Ac and Leg5AmNMe7Ac). Targeted metabolomic analyses of isogenic mutants established a role for the ptmA-F genes and defined two new ptm genes in this locus as legionaminic acid biosynthetic enzymes. This is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein

  metabolism, glycan, legionaminic acid, modeling, Flagellin, Campylobacter coli

  NCBI PubMed ID: 17371878
  Journal NLM ID: 2985121R
  Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
  Correspondence: susan.logan@nrc-cnrc.gc.ca; evelyn.soo@nrc-cnrc.gc.ca
  Institutions: National Research Council, Institute for Biological Sciences, Ottawa, Ontario, Canada
  Methods: 13C NMR, 1H NMR, NMR-2D, MD simulations, NMR-1D, genetic methods, biochemical methods, biosynthetic methods, CE-ESI-MS, LC-MS
  
   
  
  Campylobacter coli VC167
  CSDB ID 21877 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-pseudaminic acid
Compound class: nucleoside monophosphate sugar
Contained glycoepitopes: IEDB_141494,IEDB_167475,IEDB_838988

• Article ID: 3403
  Guerry P, Ewing CP, Schoenhofen IC, Logan SM "Protein glycosylation in Campylobacter jejuni: partial suppression of pglF by mutation of pseC" - Journal of Bacteriology 189(18) (2007) 6731-6733
  

  Campylobacter jejuni has systems for N- and O-linked protein glycosylation. Although biochemical evidence demonstrated that a pseC mutant in the O-linked pathway accumulated the product of pglF in the N-linked pathway, analyses of transformation frequencies and glycosylation statuses of N-glycosylated proteins indicated a partial suppression of pglF by pseC

  biosynthesis, Bacterial Proteins, Campylobacter jejuni, glycosylation

  NCBI PubMed ID: 17631632
  Journal NLM ID: 2985120R
  Publisher: American Society for Microbiology
  Correspondence: guerryp@nmrc.navy.mil
  Institutions: Enteric Diseases Department, Naval Medical Research Center, 503 Robert Grant Ave., Silver Spring, MD 20910, USA
  Methods: PCR, serological methods, genetic methods
  
   
  
  Campylobacter jejuni
  CSDB ID 21965 (all data & tools)
• Article ID: 3823
  Brisson JR, Vinogradov E, McNally DJ, Khieu NH, Schoenhofen IC, Logan SM, Jarrell H "The application of NMR spectroscopy to functional glycomics" - Book: Methods in Molecular Biology (2010) Vol. 600, 155-173
  

  Glycomics which is the study of saccharides and genes responsible for their formation requires the continuous development of rapid and sensitive methods for the identification of glycan structures. It involves glycoanalysis which relies upon the development of methods for determining the structure and interactions of carbohydrates. For the application of functional glycomics to microbial virulence, carbohydrates and their associated metabolic and carbohydrate processing enzymes and respective genes can be identified and exploited as targets for drug discovery, glyco-engineering, vaccine design, and detection and diagnosis of diseases. Glycomics also encompasses the detailed understanding of carbohydrate-protein interactions and this knowledge can be applied to research efforts focused toward the development of vaccines and immunological therapies to alleviate infectious diseases.

  NMR, polysaccharides, structural analysis, molecular modeling, HR-MAS, Glycomics, glycans, glycoanalysis, protein–carbohydrate interactions

  NCBI PubMed ID: 19882127
  Publisher: Totowa, NJ: Humana Press
  Editors: Holst O, Walker JM, Beck A
  Institutions: Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada
  Methods: NMR
  
   
  
  Helicobacter pylori, Campylobacter jejuni
  CSDB ID 25548 (all data & tools)
• Article ID: 4994
  Friedrich V, Janesch B, Windwarder M, Maresch D, Braun ML, Megson ZA, Vinogradov E, Goneau MF, Sharma A, Altmann F, Messner P, Schoenhofen IC, Schäffer C "Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications" - Glycobiology 7(4) (2017) 342-357
  

  Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium's cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead. In contrast to the NulO in ATCC 43037, which has been previously identified as a Pse derivative (5-N-acetimidoyl-7-N-glyceroyl-3,5,7,9-tetradeoxy-l-glycero-l-manno-NulO), glycan analysis of strain UB4 performed in this study indicated a 350-Da, possibly N-glycolyl Leg (3,5,7,9-tetradeoxy-d-glycero-d-galacto-NulO) derivative with unknown C5,7 N-acyl moieties. We have expressed, purified and characterized enzymes of both NulO pathways to confirm these genes' functions. Using capillary electrophoresis (CE), CE-mass spectrometry and NMR spectroscopy, our studies revealed that Pse biosynthesis in ATCC 43037 essentially follows the UDP-sugar route described in Helicobacter pylori, while the pathway in strain FDC 92A2 corresponds to Leg biosynthesis in Campylobacter jejuni involving GDP-sugar intermediates. To demonstrate that the NulO biosynthesis enzymes are functional in vivo, we created knockout mutants resulting in glycans lacking the respective NulO. Compared to the wild-type strains, the mutants exhibited significantly reduced biofilm formation on mucin-coated surfaces, suggestive of their involvement in host-pathogen interactions or host survival. This study contributes to understanding possible biological roles of bacterial NulOs.

  Campylobacter jejuni, Helicobacter, Biofilm, biosynthesis pathway, pseudaminic and legionaminic acid, bacterium

  NCBI PubMed ID: 27986835
  Publication DOI: 10.1093/glycob/cww129
  Journal NLM ID: 9104124
  Publisher: IRL Press at Oxford University Press
  Correspondence: christina.schaeffer@boku.ac.at; Ian.Schoenhofen@nrc-cnrc.gc.ca
  Institutions: Department of NanoBiotechnology, NanoGlycobiology Unit, Universität für Bodenkultur Wien, Muthgasse 11, A-1190 Vienna, Austria, Department of Chemistry, Universität für Bodenkultur Wien, Muthgasse 18, A-1190 Vienna, Austria, National Research Council, Human Health Therapeutics Portfolio, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6, Department of Oral Biology, School of Dental Medicine, University at Buffalo, 311 Foster Hall, 3435 Main St. Buffalo, New York 14214, USA
  Methods: 13C NMR, 1H NMR, PCR, SDS-PAGE, DNA techniques, b-elimination, genetic methods, biochemical methods, CE, CE-MS, bioinformatic analysis, LC-ESI-MS/MS, biofilm assays
  
   
  
  Tannerella forsythia ATCC 43037
  CSDB ID 12084 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-sialic acid
Contained glycoepitopes: IEDB_136794,IEDB_141494,IEDB_146100,IEDB_149174,IEDB_167475,SB_170,SB_171,SB_172,SB_84

• Article ID: 3474
  Glaze PA, Watson DC, Young NM, Tanner ME "Biosynthesis of CMP-N,N'-diacetyllegionaminic acid from UDP-N,N'-diacetylbacillosamine in Legionella pneumophila" - Biochemistry 47(10) (2008) 3272-3282
  

  Legionaminic acid is a nine-carbon α-keto acid that is similar in structure to other members of the sialic acid family that includes neuraminic acid and pseudaminic acid. It is found as a component of the lipopolysaccharide in several bacterial species and is perhaps best known for its presence in the O-antigen of the causative agent of Legionnaires' disease, Legionella pneumophila. In this work, the enzymes responsible for the biosynthesis and activation of N, N'-diacetyllegionaminic acid are identified for the first time. A cluster of three L. pneumophila genes bearing homology to known sialic acid biosynthetic genes (neuA,B,C) were cloned and overexpressed in Escherichia coli. The NeuC homologue was found to be a hydrolyzing UDP- N, N'-diacetylbacillosamine 2-epimerase that converts UDP- N, N'-diacetylbacillosamine into 2,4-diacetamido-2,4,6-trideoxymannose and UDP. Stereochemical and isotopic labeling studies showed that the enzyme utilizes a mechanism involving an initial anti elimination of UDP to form a glycal intermediate and a subsequent syn addition of water to generate product. This is similar to the hydrolyzing UDP- N-acetylglucosamine 2-epimerase (NeuC) of sialic acid biosynthesis, but the L. pneumophila enzyme would not accept UDP-GlcNAc as an alternate substrate. The NeuB homologue was found to be a N, N'-diacetyllegionaminic acid synthase that condenses 2,4-diacetamido-2,4,6-trideoxymannose with phosphoenolpyruvate (PEP), although the in vitro activity of the recombinant enzyme (isolated as a MalE fusion protein) was very low. The synthase activity was dependent on the presence of a divalent metal ion, and the reaction proceeded via a C-O bond cleavage process, similar to the reactions catalyzed by the sialic acid and pseudaminic acid synthases. Finally, the NeuA homologue was shown to possess the CMP- N, N'-diacetyllegionaminic acid synthetase activity that generates the activated form of legionaminic acid used in lipopolysaccharide biosynthesis. Together, the three enzymes constitute a pathway that converts a UDP-linked bacillosamine derivative into a CMP-linked legionaminic acid derivative

  O-antigen, lipopolysaccharide biosynthesis, pseudaminic acid, neuraminic acid, Legionella pneumophila, legionaminic acid, Legionella, bacillosamine

  NCBI PubMed ID: 18275154
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Correspondence: mtanner@chem.ubc.ca
  Institutions: Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1
  Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, 31P NMR, genetic methods, biochemical methods
  
   
  
  Legionella pneumophila
  CSDB ID 22962 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CMP-pseudaminic acid, CMP-Pse5Ac7Ac
Contained glycoepitopes: IEDB_141494,IEDB_167475,IEDB_838988

• Article ID: 3505
  Liu B, Knirel YA, Feng L, Perepelov AV, Senchenkova SN, Wang Q, Reeves P, Wang L "Structure and genetics of Shigella O antigens" - FEMS Microbiology Reviews 32(4) (2008) 627-653
  

  This review covers the O antigens of the 46 serotypes of Shigella, but those of most Shigella flexneri are variants of one basic structure, leaving 34 Shigella distinct O antigens to review, together with their gene clusters. Several of the structures and gene clusters are reported for the first time and this is the first such group for which structures and DNA sequences have been determined for all O antigens. Shigella strains are in effect Escherichia coli with a specific mode of pathogenicity, and 18 of the 34 O antigens are also found in traditional E. coli. Three are very similar to E. coli O antigens and 13 are unique to Shigella strains. The O antigen of Shigella sonnei is quite atypical for E. coli and is thought to have transferred from Plesiomonas. The other 12 O antigens unique to Shigella strains have structures that are typical of E. coli, but there are considerably more anomalies in their gene clusters, probably reflecting recent modification of the structures. Having the complete set of structures and genes opens the way for experimental studies on the role of this diversity in pathogenicity.

  structure, O antigen, Shigella, O antigen gene cluster, O antigen diversity

  NCBI PubMed ID: 18422615
  Publication DOI: 10.1111/j.1574-6976.2008.00114.x
  Journal NLM ID: 8902526
  Publisher: Oxford University Press
  Correspondence: wanglei@nankai.edu.cn
  Institutions: TEDA School of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, China, TEDA School of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, China.
  Methods: 13C NMR, 1H NMR, NMR-2D, sugar analysis, ESI-MS, serological methods, genetic methods, biochemical methods
  
   
  
  Shigella
  CSDB ID 23115 (all data & tools)
• Article ID: 3515
  McNally DJ, Schoenhofen IC, Houliston RS, Khieu NH, Whitfield DM, Logan SM, Jarrell HC, Brisson JR "CMP-Pseudaminic Acid is a Natural Potent Inhibitor of PseB, the First Enzyme of the Pseudaminic Acid Pathway in Campylobacter jejuni and Helicobacter pylori" - ChemMedChem 3(1) (2008) 55-59
  

  No abstract available

  Research, acid, Campylobacter, Campylobacter jejuni, NMR spectroscopy, pseudaminic acid, biological, Helicobacter pylori, enzyme, natural, pathway, Helicobacter, inhibitors, STD, STD NMR spectroscopy, potent

  NCBI PubMed ID: 17893902
  Journal NLM ID: 101259013
  Publisher: Wiley-VCH
  Correspondence: david.mcnally@nrc-cnrc.gc.ca
  Institutions: National Research Council of Canada-Institute for Biological Sciences, Ottawa ON, K1A 0R6, Canada, Fax: (+1) 613-952-9092
  Methods: 13C NMR, 1H NMR, NMR-2D, sugar analysis, NMR-1D, genetic methods, biochemical methods, STD NMR
  
   
  
  Campylobacter jejuni, Helicobacter pylori
  CSDB ID 23138 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CDP-ribitol
Contained glycoepitopes: IEDB_114703,IEDB_141494,IEDB_167475

• Article ID: 3663
  Baur S, Marles-Wright J, Buckenmaier S, Lewis RJ, Vollmer W "Synthesis of CDP-activated ribitol for teichoic acid precursors in Streptococcus pneumoniae" - Journal of Bacteriology 191(4) (2009) 1200-1210
  

  Streptococcus pneumoniae has unusually complex cell wall teichoic acid and lipoteichoic acid, both of which contain a ribitol phosphate moiety. The lic region of the pneumococcal genome contains genes for the uptake and activation of choline, the attachment of phosphorylcholine to teichoic acid precursors, and the transport of these precursors across the cytoplasmic membrane. The role of two other, so far uncharacterized, genes, spr1148 and spr1149, in the lic region was determined. TarJ (spr1148) encodes an NADPH-dependent alcohol dehydrogenase for the synthesis of ribitol 5-phosphate from ribulose 5-phosphate. TarI (spr1149) encodes a cytidylyl transferase for the synthesis of cytidine 5'-diphosphate (CDP)-ribitol from ribitol 5-phosphate and cytidine 5'-triphosphate. We also present the crystal structure of TarI with and without bound CDP, and the structures present a rationale for the substrate specificity of this key enzyme. No transformants were obtained with insertion plasmids designed to interrupt the tarIJ genes, indicating that their function could be essential for cell growth. CDP-activated ribitol is a precursor for the synthesis of pneumococcal teichoic acids and some of the capsular polysaccharides. Thus, all eight genes in the lic region have a role in teichoic acid synthesis.

  synthesis, Streptococcus pneumoniae, transferase, cell wall, crystal structure, teichoic acids

  NCBI PubMed ID: 19074383
  Journal NLM ID: 2985120R
  Publisher: American Society for Microbiology
  Correspondence: W.Vollmer@ncl.ac.uk
  Institutions: Institute for Cell and Molecular Biosciences, Newcastle University, Framlington Place, Newcastle upon Tyne NE2 4HH, United Kingdom
  Methods: MALDI-TOF MS, serological methods, genetic methods, biochemical methods, crystallization
  
   
  
  Streptococcus pneumoniae
  CSDB ID 23652 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CDP-D-Glcp
Contained glycoepitopes: IEDB_141494,IEDB_142488,IEDB_144998,IEDB_145000,IEDB_145002,IEDB_146664,IEDB_167475,IEDB_983931,SB_192

• Article ID: 3679
  Cunneen MM, De Castro C, Kenyon J, Parrilli M, Reeves PR, Molinaro A, Holst O, Skurnik M "The O-specific polysaccharide structure and biosynthetic gene cluster of Yersinia pseudotuberculosis serotype O:11" - Carbohydrate Research 344(12) (2009) 1533-1540
  

  In the Yersinia pseudotuberculosis serotyping scheme, 21 serotypes are present originating from about 30 different O-factors distributed within the species. With regard to the chemical structures of lipopolysaccharides (LPSs) and the genetic basis of their biosynthesis, a number, but not all, of Y. pseudotuberculosis strains representing different serotypes have been investigated. In order to present an overall picture of the relationship between genetics and structures, we have been working on the genetics and structures of various Y. pseudotuberculosis O-specific polysaccharides (OPSs). Here, we present a structural and genetic analysis of the Y. pseudotuberculosis serotype O:11 OPS. Our results showed that this OPS structure has the same backbone as that of Y. pseudotuberculosis O:1b, but with a 6d-l-Altf side-branch instead of Parf. The 3' end of the gene cluster is the same as that for O:1b and has the genes for synthesis of the backbone and for processing the completed repeat unit. The 5' end has genes for synthesis of 6d-l-Altf and its transfer to the repeating unit backbone. The pathway for the synthesis of the 6d-l-Altf appears to be different from that for 6d-l-Altp in Y. enterocolitica O:3. The chemical structure of the O:11 repeating unit is

  structure, O-specific polysaccharide, Yersinia pseudotuberculosis, O-specific polysaccharides, 6-Deoxy-L-altrofuranose, biosynthetic gene cluster

  NCBI PubMed ID: 19505680
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: oholst@fz-borstel.de (O. Holst)
  Institutions: Division of Microbiology, School of Molecular and Microbial Biosciences, University of Sydney, Australia
  Methods: 13C NMR, 1H NMR, NMR-2D, methylation, GC-MS, sugar analysis, NMR-1D, genetic methods
  
   
  
  Yersinia pseudotuberculosis O11
  CSDB ID 23984 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CDP-6-deoxy-D-xylo-hex-4-ulopyranose
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3679
  Cunneen MM, De Castro C, Kenyon J, Parrilli M, Reeves PR, Molinaro A, Holst O, Skurnik M "The O-specific polysaccharide structure and biosynthetic gene cluster of Yersinia pseudotuberculosis serotype O:11" - Carbohydrate Research 344(12) (2009) 1533-1540
  

  In the Yersinia pseudotuberculosis serotyping scheme, 21 serotypes are present originating from about 30 different O-factors distributed within the species. With regard to the chemical structures of lipopolysaccharides (LPSs) and the genetic basis of their biosynthesis, a number, but not all, of Y. pseudotuberculosis strains representing different serotypes have been investigated. In order to present an overall picture of the relationship between genetics and structures, we have been working on the genetics and structures of various Y. pseudotuberculosis O-specific polysaccharides (OPSs). Here, we present a structural and genetic analysis of the Y. pseudotuberculosis serotype O:11 OPS. Our results showed that this OPS structure has the same backbone as that of Y. pseudotuberculosis O:1b, but with a 6d-l-Altf side-branch instead of Parf. The 3' end of the gene cluster is the same as that for O:1b and has the genes for synthesis of the backbone and for processing the completed repeat unit. The 5' end has genes for synthesis of 6d-l-Altf and its transfer to the repeating unit backbone. The pathway for the synthesis of the 6d-l-Altf appears to be different from that for 6d-l-Altp in Y. enterocolitica O:3. The chemical structure of the O:11 repeating unit is

  structure, O-specific polysaccharide, Yersinia pseudotuberculosis, O-specific polysaccharides, 6-Deoxy-L-altrofuranose, biosynthetic gene cluster

  NCBI PubMed ID: 19505680
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: oholst@fz-borstel.de (O. Holst)
  Institutions: Division of Microbiology, School of Molecular and Microbial Biosciences, University of Sydney, Australia
  Methods: 13C NMR, 1H NMR, NMR-2D, methylation, GC-MS, sugar analysis, NMR-1D, genetic methods
  
   
  
  Yersinia pseudotuberculosis O11
  CSDB ID 23985 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: CDP-6-deoxy-L-arabino-hex-4-ulopyranose
Contained glycoepitopes: IEDB_141494,IEDB_167475

• Article ID: 3679
  Cunneen MM, De Castro C, Kenyon J, Parrilli M, Reeves PR, Molinaro A, Holst O, Skurnik M "The O-specific polysaccharide structure and biosynthetic gene cluster of Yersinia pseudotuberculosis serotype O:11" - Carbohydrate Research 344(12) (2009) 1533-1540
  

  In the Yersinia pseudotuberculosis serotyping scheme, 21 serotypes are present originating from about 30 different O-factors distributed within the species. With regard to the chemical structures of lipopolysaccharides (LPSs) and the genetic basis of their biosynthesis, a number, but not all, of Y. pseudotuberculosis strains representing different serotypes have been investigated. In order to present an overall picture of the relationship between genetics and structures, we have been working on the genetics and structures of various Y. pseudotuberculosis O-specific polysaccharides (OPSs). Here, we present a structural and genetic analysis of the Y. pseudotuberculosis serotype O:11 OPS. Our results showed that this OPS structure has the same backbone as that of Y. pseudotuberculosis O:1b, but with a 6d-l-Altf side-branch instead of Parf. The 3' end of the gene cluster is the same as that for O:1b and has the genes for synthesis of the backbone and for processing the completed repeat unit. The 5' end has genes for synthesis of 6d-l-Altf and its transfer to the repeating unit backbone. The pathway for the synthesis of the 6d-l-Altf appears to be different from that for 6d-l-Altp in Y. enterocolitica O:3. The chemical structure of the O:11 repeating unit is

  structure, O-specific polysaccharide, Yersinia pseudotuberculosis, O-specific polysaccharides, 6-Deoxy-L-altrofuranose, biosynthetic gene cluster

  NCBI PubMed ID: 19505680
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: oholst@fz-borstel.de (O. Holst)
  Institutions: Division of Microbiology, School of Molecular and Microbial Biosciences, University of Sydney, Australia
  Methods: 13C NMR, 1H NMR, NMR-2D, methylation, GC-MS, sugar analysis, NMR-1D, genetic methods
  
   
  
  Yersinia pseudotuberculosis O11
  CSDB ID 23986 (all data & tools)