A FLUORESCENT ACTIVITY ASSAY FOR HIGH-THROUGHPUT SCREENING OF O- GlcNAc TRANSFERASE INHIBITORS

摘要

The addition of O-linked N-acetylglucosamine (O-GlcNAc) to serine and threonine residues of proteins is a dynamic and ubiquitous post-translational modification in mammalian cells [1]. This modification is mediated by only two enzymes; O-GlcNAc transferase (OGT), which catalyzes the addition of the O-GlcNAc unit, and O-GlcNAcase (OGA), which catalyzes hydrolysis of the carbohydrate unit from the protein sidechain. OGT is emerging as a major regulator of cell and organism physiology, regulating central processes such as transcription [2]. Further, dysregulation of the O-GlcNAc modification has been implicated in numerous disorders including Alzheimer's Disease and cancers [3,4]. Unfortunately, there are limited chemical biology tools available to study this modification and its fundamental roles in health and disease. Inhibitors of OGT, in particular, are expected to have great potential both for studying the role OGT in living systems as well as for the treatment of diseases which are linked to excessive OGT activity.