Literature DB >> 17346239

Identification and biochemical characterization of two novel UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronic acid 2-epimerases from respiratory pathogens.

Erin L Westman1, David J McNally, Martin Rejzek, Wayne L Miller, Vellupillai Sri Kannathasan, Andrew Preston, Duncan J Maskell, Robert A Field, Jean-Robert Brisson, Joseph S Lam.   

Abstract

The heteropolymeric O-antigen of the lipopolysaccharide from Pseudomonas aeruginosa serogroup O5 as well as the band-A trisaccharide from Bordetella pertussis contain the di-N-acetylated mannosaminuronic acid derivative, beta-D-ManNAc3NAcA (2,3-diacetamido-2,3-dideoxy-beta-D-mannuronic acid). The biosynthesis of the precursor for this sugar is proposed to require five steps, through which UDP-alpha-D-GlcNAc (UDP-N-acetyl-alpha-D-glucosamine) is converted via four steps into UDP-alpha-D-GlcNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronic acid), and this intermediate compound is then epimerized by WbpI (P. aeruginosa), or by its orthologue, WlbD (B. pertussis), to form UDP-alpha-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-mannuronic acid). UDP-alpha-D-GlcNAc3NAcA, the proposed substrate for WbpI and WlbD, was obtained through chemical synthesis. His6-WbpI and His6-WlbD were overexpressed and then purified by affinity chromatography using FPLC. Capillary electrophoresis was used to analyse reactions with each enzyme, and revealed that both enzymes used UDP-alpha-D-GlcNAc3NAcA as a substrate, and reacted optimally in sodium phosphate buffer (pH 6.0). Neither enzyme utilized UDP-alpha-D-GlcNAc, UDP-alpha-D-GlcNAcA (UDP-2-acetamido-2,3-dideoxy-alpha-D-glucuronic acid) or UDP-alpha-D-GlcNAc3NAc (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucose) as substrates. His6-WbpI or His6-WlbD reactions with UDP-alpha-D-GlcNAc3NAcA produce a novel peak with an identical retention time, as shown by capillary electrophoresis. To unambiguously characterize the reaction product, enzyme-substrate reactions were allowed to proceed directly in the NMR tube and conversion of substrate into product was monitored over time through the acquisition of a proton spectrum at regular intervals. Data collected from one- and two-dimensional NMR experiments showed that His6-WbpI catalysed the 2-epimerization of UDP-alpha-D-GlcNAc3NAcA, converting it into UDP-alpha-D-ManNAc3NAcA. Collectively, these results provide evidence that WbpI and WlbD are UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronic acid 2-epimerases.

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Year:  2007        PMID： 17346239      PMCID： PMC1925246          DOI： 10.1042/BJ20070017

Source DB:  PubMed          Journal:  Biochem J        ISSN： 0264-6021            Impact factor:   3.857


  26 in total

1.  Molecular characterization of the Pseudomonas aeruginosa serotype O5 (PAO1) B-band lipopolysaccharide gene cluster.

Authors:  L L Burrows; D F Charter; J S Lam
Journal:  Mol Microbiol       Date:  1996-11       Impact factor: 3.501

Review 2.  Solution nuclear magnetic resonance spectroscopy techniques for probing intermolecular interactions.

Authors:  Maurizio Pellecchia
Journal:  Chem Biol       Date:  2005-09

3.  Structural studies of the lipopolysaccharide O-antigen and capsular polysaccharide of Vibrio anguillarum serotype O:2.

Authors:  I Sadovskaya; J R Brisson; E Altman; L M Mutharia
Journal:  Carbohydr Res       Date:  1996-03-22       Impact factor: 2.104

4.  Recognition of a bacterial adhesion by an integrin: macrophage CR3 (alpha M beta 2, CD11b/CD18) binds filamentous hemagglutinin of Bordetella pertussis.

Authors:  D Relman; E Tuomanen; S Falkow; D T Golenbock; K Saukkonen; S D Wright
Journal:  Cell       Date:  1990-06-29       Impact factor: 41.582

5.  Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7.

Authors:  Y A Knirel; N A Paramonov; E V Vinogradov; A S Shashkov; B A Dmitriev; N K Kochetkov; E V Kholodkova; E S Stanislavsky
Journal:  Eur J Biochem       Date:  1987-09-15

6.  Staphylococcus aureus cap5P encodes a UDP-N-acetylglucosamine 2-epimerase with functional redundancy.

Authors:  K B Kiser; N Bhasin; L Deng; J C Lee
Journal:  J Bacteriol       Date:  1999-08       Impact factor: 3.490

7.  Mapping the active site of the Haemophilus influenzae methionyl-tRNA formyltransferase: residues important for catalysis and tRNA binding.

Authors:  D T Newton; D Mangroo
Journal:  Biochem J       Date:  1999-04-01       Impact factor: 3.857

8.  Evidence that WbpD is an N-acetyltransferase belonging to the hexapeptide acyltransferase superfamily and an important protein for O-antigen biosynthesis in Pseudomonas aeruginosa PAO1.

Authors:  Cory Q Wenzel; Craig Daniels; Robert A B Keates; Dyanne Brewer; Joseph S Lam
Journal:  Mol Microbiol       Date:  2005-09       Impact factor: 3.501

9.  The identification, cloning and mutagenesis of a genetic locus required for lipopolysaccharide biosynthesis in Bordetella pertussis.

Authors:  A Allen; D Maskell
Journal:  Mol Microbiol       Date:  1996-01       Impact factor: 3.501

10.  Characterization of lipopolysaccharide-deficient mutants of Pseudomonas aeruginosa derived from serotypes O3, O5, and O6.

Authors:  T Dasgupta; T R de Kievit; H Masoud; E Altman; J C Richards; I Sadovskaya; D P Speert; J S Lam
Journal:  Infect Immun       Date:  1994-03       Impact factor: 3.441
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  8 in total

1.  Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa.

Authors:  Erin L Westman; David J McNally; Armen Charchoglyan; Dyanne Brewer; Robert A Field; Joseph S Lam
Journal:  J Biol Chem       Date:  2009-03-12       Impact factor: 5.157

2.  Biosynthesis of a rare di-N-acetylated sugar in the lipopolysaccharides of both Pseudomonas aeruginosa and Bordetella pertussis occurs via an identical scheme despite different gene clusters.

Authors:  Erin L Westman; Andrew Preston; Robert A Field; Joseph S Lam
Journal:  J Bacteriol       Date:  2008-07-11       Impact factor: 3.490

3.  Biosynthesis of UDP-GlcNAc(3NAc)A by WbpB, WbpE, and WbpD: enzymes in the Wbp pathway responsible for O-antigen assembly in Pseudomonas aeruginosa PAO1.

Authors:  Angelyn Larkin; Barbara Imperiali
Journal:  Biochemistry       Date:  2009-06-16       Impact factor: 3.162

4.  Enzymatic analysis of uridine diphosphate N-acetyl-D-glucosamine.

Authors:  Seema C Namboori; David E Graham
Journal:  Anal Biochem       Date:  2008-06-27       Impact factor: 3.365

5.  Predicting protein function from structure--the roles of short-chain dehydrogenase/reductase enzymes in Bordetella O-antigen biosynthesis.

Authors:  Jerry D King; Nicholas J Harmer; Andrew Preston; Colin M Palmer; Martin Rejzek; Robert A Field; Tom L Blundell; Duncan J Maskell
Journal:  J Mol Biol       Date:  2007-09-26       Impact factor: 5.469

6.  Involvement of the Wbp pathway in the biosynthesis of Porphyromonas gingivalis lipopolysaccharide with anionic polysaccharide.

Authors:  Mikio Shoji; Keiko Sato; Hideharu Yukitake; Mariko Naito; Koji Nakayama
Journal:  Sci Rep       Date:  2014-05-23       Impact factor: 4.379

7.  Functional characterization of the rice UDP-glucose 4-epimerase 1, OsUGE1: a potential role in cell wall carbohydrate partitioning during limiting nitrogen conditions.

Authors:  David R Guevara; Ashraf El-Kereamy; Mahmoud W Yaish; Yong Mei-Bi; Steven J Rothstein
Journal:  PLoS One       Date:  2014-05-01       Impact factor: 3.240

8.  Characterization of Structure and Antioxidant Activity of Polysaccharides From Sesame Seed Hull.

Authors:  Run-Yang Zhang; Jing-Hao Gao; Yi-Lin Shi; Yi-Fei Lan; Hua-Min Liu; Wen-Xue Zhu; Xue-De Wang
Journal:  Front Nutr       Date:  2022-06-21
  8 in total

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