Tools for probing and perturbing O-GlcNAc in cells and in vivo.

Intracellular glycosylation of nuclear and cytoplasmic proteins involves the addition of N-acetylglucosamine (O-GlcNAc) to serine and threonine residues. This dynamic modification occurs on hundreds of proteins and is involved in various essential biological processes. Because O-GlcNAc is substoichiometric and labile, identifying proteins and sites of modification has been challenging and generally requires proteome enrichment. Here we review recent advances on the implementation of chemical tools to perturb, to detect, and to map O-GlcNAc in living systems. Metabolic and chemoenzymatic labels along with bioorthogonal reactions and quantitative proteomics are enabling investigation of the role of O-GlcNAc in various processes including transcriptional regulation, neurodegeneration, and cell signaling.