Click chemistry-based universal enrichment columns for global identification of azide-modified proteins

摘要

We show the efficient capture of metabolically labeled O-GlcNAcylated proteins from Jurkat cell lysates onto a click chemistry based resin with greater than 63percent enrichment. The covalent nature of the attachment of these proteins to the clickable resin allows for highly stringent washing conditions to eliminate the non-specific background protein binding, which is a major shortcoming of biotin-based enrichment columns. Protease digestion of the resin-bound O-GIcNAcylated proteins generated highly enriched peptide pools that have greater than 85percent purity. The peptides are compatible with mass spectrometry based proteomic studies for global identification of PTM proteins.