ArticleBiochemistry2006
Vi antigen biosynthesis in Salmonella typhi: characterization of UDP-N-acetylglucosamine C-6 dehydrogenase (TviB) and UDP-N-acetylglucosaminuronic acid C-4 epimerase (TviC).
Article in Biochemistry, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 30 citations in OpenAlex.
- Protein-centric mechanisms of typhoidalFrontiers in cellular and infection microbiology · 2026Review
- Genetic heterogeneity in theMicrobial genomics · 2024Article
- Single missense mutations in Vi capsule synthesis genes confer hypervirulence to Salmonella Typhi.Nature communications · 2024Article
- Characterization ofmBio · 2024Article
- Enteric Fever Diagnosis: Current Challenges and Future Directions.Pathogens (Basel, Switzerland) · 2021Review
- A whole-genome screen identifies Salmonella enterica serovar Typhi genes involved in fluoroquinolone susceptibility.The Journal of antimicrobial chemotherapy · 2020Article
- The UDP-GalNAcA biosynthesis genes gna-gne2 are required to maintain cell envelope integrity and in vivo fitness in multi-drug resistant Acinetobacter baumannii.Molecular microbiology · 2020Article
- Sewage effluent from an Indian hospital harbors novel carbapenemases and integron-borne antibiotic resistance genes.Microbiome · 2019Article
- Why Is Eradicating Typhoid Fever So Challenging: Implications for Vaccine and Therapeutic Design.Vaccines · 2018Review
- Survival of the Fittest: How Bacterial Pathogens Utilize Bile To Enhance Infection.Clinical microbiology reviews · 2016Review
- Characterization and Comparative Overview of Complete Sequences of the First Plasmids ofFrontiers in microbiology · 2016Article
- Comparative genomic and transcriptomic analysis of wangiella dermatitidis, a major cause of phaeohyphomycosis and a model black yeast human pathogen.G3 (Bethesda, Md.) · 2014Article
- Variation in the complex carbohydrate biosynthesis loci of Acinetobacter baumannii genomes.PloS one · 2013Article
- Molecular characterization of the viaB locus encoding the biosynthetic machinery for Vi capsule formation in Salmonella Typhi.PloS one · 2012Article
- Transcriptional regulation of the Escherichia coli gene rraB, encoding a protein inhibitor of RNase E.Journal of bacteriology · 2009Article
- Identification of the UDP-N-acetylglucosamine 4-epimerase involved in exosporium protein glycosylation in Bacillus anthracis.Journal of bacteriology · 2009Article
- Regulation of Vi capsular polysaccharide synthesis in Salmonella enterica serotype Typhi.Journal of infection in developing countries · 2008Article
- Enzymatic analysis of uridine diphosphate N-acetyl-D-glucosamine.Analytical biochemistry · 2008Article
- Acetamido sugar biosynthesis in the Euryarchaea.Journal of bacteriology · 2008Article
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4 authors at 1 institution in 1 country.
Funding
Abstract
Vi antigen, the virulence factor of Salmonella typhi, has been used clinically as a molecular vaccine. TviB and TviC are two enzymes involved in the formation of Vi antigen, a linear polymer consisting of alpha-1,4-linked N-acetylgalactosaminuronate. Protein sequence analysis suggests that TviB is a dehydrogenase and TviC is an epimerase. Both enzymes are expected to be NAD(+) dependent. In order to verify their functions, TviB and TviC were cloned, expressed in Escherichia coli, and characterized. The C-terminal His(6)-tagged TviB protein, purified from soluble cell fractions in the presence of 10 mM DTT, shows UDP-N-acetylglucosamine 6-dehydrogenase activity and is capable of catalyzing the conversion of UDP-N-acetylglucosamine (UDP-GlcNAc) to UDP-N-acetylglucosaminuronic acid (UDP-GlcNAcA) with a k(cat) value of 15.5 +/- 1.0 min(-)(1). The K(m) values of TviB for UDP-GlcNAc and NAD(+) are 77 +/- 9 microM and 276 +/- 52 microM, respectively. TviC, purified as C-terminal hexahistidine-tagged protein, shows UDP-GlcNAcA 4-epimerase and UDP-N-acetylgalactosamine (UDP-GalNAc) 4-epimerase activities. The K(m) values of TviC for UDP-GlcNAcA and UDP-N-acetylgalactosaminuronic acid (UDP-GalNAcA) are 20 +/- 1 microM and 42 +/- 2 microM, respectively. The k(cat) value for the conversion of UDP-GlcNAcA to UDP-GalNAcA is 56.8 +/- 0.5 min(-)(1), while that for the reverse reaction is 39.1 +/- 0.6 min(-)(1). These results show that the biosynthesis of Vi antigen is initiated by the TviB-catalyzed oxidation of UDP-GlcNAc to UDP-GalNAc, followed by the TviC-catalyzed epimerization at C-4 to form UDP-GalNAcA, which serves as the building block for the formation of Vi polymer. These results set the stage for future in vitro biosynthesis of Vi antigen. These enzymes may also be drug targets to inhibit Vi antigen production.
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