Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
The structure was elucidated in this paperNCBI PubMed ID: 18179256Publication DOI: 10.1021/bi7020712Journal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: wang.892
osu.edu
Institutions: Department of Biochemistry, Department of Chemistry, and The Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio 43210
T-Antigen (Gal-β1,3-GalNAc-α-O-Ser/Thr) is an important precursor of mucin-type O-glycans. T-Antigen is found to be closely associated with cancer progression and metastasis and has been used to develop carbohydrate-based anticancer vaccines. Enzymatic synthesis of T-antigen disaccharides have relied on the use of β-1,3-galactosyltransferases recently cloned and characterized from several eukaryotic organisms. However, its application is limited by the difficulty of obtaining homogeneous enzymes and the strict substrate specificity of enzymes. Recently, a number of bacteria have been found to express carbohydrate structures that mimic host glycans. The corresponding glycosyltransferases have been exploited in the facile synthesis of a number of clinically important glycoconjugate mimics. In this study, we biochemically characterized a bacterial β-1,3-galactosyltransferase (WbiP) from Escherichia coli O127, which expresses a T-antigen mimic in the lipopolysaccharide (LPS) structure. Substrate study showed that WbiP could readily glycosylate a series of N-acetylgalactosamine (GalNAc) analogues with alpha-substitutions at the reducing end, including glycosylated Ser and Thr (GalNAc-α-O-Ser/Thr), which illustrates the use of WbiP for the facile synthesis of T-antigens. Alignment of a group of putative bacterial β-1,3-galactosyltransferases revealed the presence of two conserved DXD motifs, possibly suggesting a different functional role of each motif. Site-directed mutagenesis, enzyme kinetics as well as UDP-bead binding assays were carried out to investigate the role of each DXD motif in WbiP. The results suggest that 88DSD90 is critical in the binding of sugar donor UDP-Gal, whereas 174DYD176 may participate in the binding of the sugar acceptor. This study expands the scope of using bacterial glycosyltransferases as tools for in vitro synthesis of glycoconjugate mimics with clinical significance
Escherichia coli, glycosyltransferases, T-Antigen, β-1, 3-Galactosyltransferase
Structure type: oligomer
Location inside paper: p. 1244, fig 2
Aglycon: 2-aminobezamide
Contained glycoepitopes: IEDB_130648,IEDB_134627,IEDB_136044,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_1391963,IEDB_141584,IEDB_141794,IEDB_143260,IEDB_190606,IEDB_885822,SB_165,SB_166,SB_187,SB_195,SB_23,SB_24,SB_7,SB_8,SB_88
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, 31P NMR, ESI-MS, MALDI-MS, NMR-1D, genetic methods, biochemical methods, HPLC
Enzymes that release or process the structure: WbiP b-1,3-galactosyltransferase
Biosynthesis and genetic data: genetic data, biochemical data
Synthetic data: enzymatic
Comments, role: The reaction product was labeled with 2-aminobezamide.
Related record ID(s): 22757, 23334, 23335, 23336
NCBI Taxonomy refs (TaxIDs): 562Reference(s) to other database(s): GTC:G00031MO, GlycomeDB:
475
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There is only one chemically distinct structure:
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