Glycobiology. 2026 Oct 9:cwag095. doi: 10.1093/glycob/cwag095. Online ahead of print.

ABSTRACT

Chemistry, through the synthesis of a radioactive sugar-donor, played a critical role in the discovery of O-GlcNAc and has been contributing indispensable tools ever since. Here, we cover where chemical methods have contributed to both the discovery and characterization of O-GlcNAc modifications. We will first discuss metabolic labeling and chemoenzymatic modification as two techniques that leverage the power of bioorthogonal chemistry to visualize, enrich, and even quantify O-GlcNAc modifications from living systems. We will then describe how solid-phase peptide synthesis can be combined with protein ligation methods as the only current pathway to homogeneously O-GlcNAc-modified proteins for downstream biological experiments. In each section, we point out the positives and considerations for each approach and provide protocols for normalization of best practices across the field.

PMID:42853187 | DOI:10.1093/glycob/cwag095