Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
NCBI PubMed ID: 24056013Publication DOI: 10.1016/j.carres.2013.08.021Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: daweizhou
nankai.edu.cn
Institutions: TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 HongDa Street, TEDA, Tianjin, China
In this study, synthetic acceptor substrate GlcNAc α-PO3-PO3-(CH2)11-O-phenyl (GlcNAc-PP-PhU) was employed in glycosyl transferase assays to characterize the WbuP galactosyltransferase activity. This activity was time- and enzyme concentration-dependent. The optimal enzyme activity was observed at pH 6.5 and 25°C. The enzyme requires Mn(2+) ions for maximal activity and detergents in the assay did not increase glycosyltransfer activity. The enzyme was shown to be specific for the UDP-Gal donor substrate. Kinetic parameters were determined for UDP-Gal, and GlcNAc-PP-PhU. The enzyme product was determined to have a ?-1,3-linkage using strategies based on exoglycosidase digestion combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) as well as collision-induced dissociation electrospray ionization ion trap multiple tandem MS (CID-ESI-IT-MS(n)). Our results conclusively demonstrate that the wbuP gene of Escherichia coli O114 encodes a UDP-Gal: GlcNAc ?-pyrophosphate-lipid ?-1,3-Gal-transferase that transfers the second sugar moiety in the assembly of the O114 repeating unit.
Escherichia coli, mass spectrometry, glycosyl transferase, Functional characterization, Escherichia coli O-antigen
Structure type: suggested polymer biological repeating unit
Location inside paper: p. 44, fig.1
The structure in this paper was incorrect:
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_136044,IEDB_137340,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_149136,IEDB_150899,IEDB_150900,IEDB_151531,IEDB_190606,SB_137,SB_165,SB_166,SB_187,SB_195,SB_29,SB_7,SB_88
Methods: PCR, GC-MS, SDS-PAGE, glycosyltransferase assays, kinetics assays, ESI-MS, MALDI-TOF MS, CID-MS/MS, biochemical methods
Enzymes that release or process the structure: WbuP b-1,3-galactosyltransferase
Synthetic data: chemoenzymatic
Related record ID(s): 9539, 20674, 22457, 25584, 28333, 108673, 122750, 137911
NCBI Taxonomy refs (TaxIDs): 562
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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