Synthesis, characterization and application of chemical tools for investigating the role of O-GlcNAc modification in the development and survival of mammalian cells

2018-10-24Functional diversity of transcribed proteins is exponentially expanded with the addition of post-translational modifications (PTMs). Understanding and characterizing the effect of these modifications on protein function and cellular health is important for developing tools for targeting their function or disfunction in cellular health and disease. Due to their post-translational and, in some cases, dynamic nature of these structural alterations, traditional structural techniques can be unreliable and difficult to reproduce. Specifically, O-GlcNAcylation describes the addition of a single monosaccharide, N-acetylglucosamine, to the hydroxyl side chains of serine or threonine residues of folded or unfolded proteins. The single enzyme responsible for installing this PTM, O-GlcNAc transferase (OGT), has an extremely broad specificity will little to no consensus substrate sequence making prediction for modification sites based on primary sequence essentially impossible. Described here is a description of technical methods used for encoding and decoding post-translational modifications, development of new chemical tools for probing post-translational glycosylation pathways and OGT, and finally the use of chemical tools for the biological study of how O-GlcNAcylation effects cell death and development