Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Host organism: Homo sapiens
Associated disease: bacterial meningitis [ICD11:
1D01.0 
];
infection due to Neisseria meningitidis [ICD11:
XN1DV ]
NCBI PubMed ID: 29228283Publication DOI: 10.1093/glycob/cwx100Journal NLM ID: 9104124Publisher: IRL Press at Oxford University Press
Correspondence: willie.vann
fda.hhs.gov
Institutions: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA, Laboratory of Bacterial Polysaccharides, FDA, Silver Spring, MD 20993, USA, Department of Chemistry, University of California, Davis, CA 95616, USA, Department of Chemistry, King's College, London SE 11DB, UK
Neisseria meningitidis Group X is an emerging cause of bacterial meningitis in Sub-Saharan Africa. The capsular polysaccharide of Group X is a homopolymer of N-acetylglucosamine α(1-4) phosphate and is a vaccine target for prevention of disease associated with this meningococcal serogroup. We have demonstrated previously that the formation of the polymer is catalyzed by a phosphotransferase which transfers N-acetylglucosamine-1-phosphate from UDP-N-acetylglucosamine to the 4-hydroxyl of the N-acetylglucosamine on the nonreducing end of the growing chain. In this study, we use substrate analogs of UDP-GlcNAc to define the enzyme/donor substrate interactions critical for catalysis. Our kinetic analysis of the phosphotransferase reaction is consistent with a sequential mechanism of substrate addition and product release. The use of novel uracil modified analogs designed by Wagner et al. enabled us to assess whether the CsxA-catalyzed reaction is consistent with a donor dependent conformational change. As expected with this model for glycosyltransferases, UDP-GlcNAc analogs with bulky uracil modifications are not substrates but are inhibitors. An analog with a smaller iodo uracil substitution is a substrate and a less potent inhibitor. Moreover, our survey of analogs with modifications on the N-acetylglucosamine residue of the sugar nucleotide donor highlights the importance of substituents at C2 and C4 of the sugar residue. The hydroxyl group at C4 and the structure of the acyl group at C2 are very important for specificity and substrate interactions during the polymerization reaction. While most analogs modified at C2 were inhibitors, acetamido analogs were also substrates suggesting the importance of the carbonyl group.
capsular polysaccharide, GlcNAc phosphotransferase, UDP-GlcNAc analogs
Structure type: homopolymer
Location inside paper: abstract
Compound class: CPS, O-polysaccharide, K-antigen, O-antigen
Contained glycoepitopes: IEDB_137340,IEDB_141807,IEDB_150077,IEDB_151531,IEDB_1711617
Methods: kinetics assays, inhibition studies, biochemical methods, HPLC, enzymatic analysis
Enzymes that release or process the structure: N-acetylglucosamine-1-phosphotransferase
NCBI Taxonomy refs (TaxIDs): 487Reference(s) to other database(s): GTC:G19079RT, GlycomeDB:
34729
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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