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Glycosyl fluorides can function as substrates for nucleotide phosphosugar-dependent glycosyltransferases

TitleGlycosyl fluorides can function as substrates for nucleotide phosphosugar-dependent glycosyltransferases
Publication TypeJournal Article
Year of Publication1999
AuthorsLougheed, B, Ly, HD, WAKARCHUK, WW, Withers, SG
JournalJOURNAL OF BIOLOGICAL CHEMISTRY
Volume274
Pagination37717-37722
Date PublishedDEC 31
ISSN0021-9258
Abstract

alpha-Galactosyl fluoride is shown to function as a substrate, in place of uridine-5'-diphosphogalactose, for the alpha-galactosyltransferase from Neisseria meningitidis. The reaction only occurs in the presence of catalytic quantities of uridine 5'-diphosphate, In the presence of galactosyl accepters, the expected oligosaccharide product is formed in essentially quantitative yields, reaction having been performed on multi-milligram scales, In the absence of a suitable acceptor, the enzyme synthesizes uridine-5'-diphosphogalactose, as demonstrated through a coupled assay in which uridine-5'-diphosphogalactose is converted to uridine-5'-diphosphoglucuronic acid with conversion of NAD to NADH. These glycosyl fluoride substrates therefore offer the potential of an inexpensive alternative donor substrate in the synthesis of oligosaccharides as well a means of generating steady state concentrations of nucleotide diphosphate sugars for in situ use by other enzymes. Further, they should prove valuable as mechanistic probes.

DOI10.1074/jbc.274.53.37717