Abstract:
:We report a straightforward enzymatic synthesis of the 4-methylumbelliferyl glycoside of a complex-type oligosaccharide substrate for core fucosylation. We demonstrate the use of this synthetic glycoconjugate in a newly developed enzyme assay to probe the activity and inhibition of fucosyltransferase VIII, which catalyzes the core fucosylation of N-glycans on eukaryotic glycoproteins. In this fucosyltransferase assay, we use the fluorogenic probe and a specific glycosidase in a sequentially coupled enzyme reaction to distinguish an unmodified 4-methylumbelliferyl oligosaccharide probe from a fucosylated probe. Our findings show that this strategy is very sensitive and specific in its detection of enzyme activity and can even be used for analyzing impure tissue lysate samples.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Soroko M,Kwan DHdoi
10.1021/acs.biochem.0c00286subject
Has Abstractpub_date
2020-06-09 00:00:00pages
2100-2110issue
22eissn
0006-2960issn
1520-4995journal_volume
59pub_type
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