Versatile O-GlcNAc transferase assay for high-throughput identification of enzyme variants, substrates, and inhibitors

Cited 17 time in scopus
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Title
Versatile O-GlcNAc transferase assay for high-throughput identification of enzyme variants, substrates, and inhibitors
Author(s)
E J Kim; L K Abramowitz; M R Bond; D C Love; D W Kang; H F Leucke; D W Kang; Jong Seog Ahn; J A Hanover
Bibliographic Citation
Bioconjugate Chemistry, vol. 25, no. 6, pp. 1025-1030
Publication Year
2014
Abstract
The dynamic glycosylation of serine/threonine residues on nucleocytoplasmic proteins with a single N-acetylglucosamine (O-GlcNAcylation) is critical for many important cellular processes. Cellular O-GlcNAc levels are highly regulated by two enzymes: O-GlcNAc transferase (OGT) is responsible for GlcNAc addition and O-GlcNAcase (OGA) is responsible for removal of the sugar. The lack of a rapid and simple method for monitoring OGT activity has impeded the efficient discovery of potent OGT inhibitors. In this study we describe a novel, single-well OGT enzyme assay that utilizes 6 × His-tagged substrates, a chemoselective chemical reaction, and unpurified OGT. The high-throughput Ni-NTA Plate OGT Assay will facilitate discovery of potent OGT-specific inhibitors on versatile substrates and the characterization of new enzyme variants.
ISSN
1043-1802
Publisher
Amer Chem Soc
DOI
http://dx.doi.org/10.1021/bc5001774
Type
Article
Appears in Collections:
Ochang Branch Institute > Chemical Biology Research Center > 1. Journal Articles
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