Bioorthogonal Chemical Reporters for Monitoring Protein Acetylation

• DOI: 10.1021/ja908871t
• Publication Date: 2010-03-01T18:00:34Z
• Authors: Yu-Ying Yang; Janice M. Ascano; Howard C. Hang
• Keywords: Bioorthogonal Chemistry

Protein acetylation is a key post-translational modification that regulates diverse biological activities in eukaryotes. Here we report bioorthogonal chemical reporters enable direct in-gel fluorescent visualization and proteome-wide identification of acetylated proteins via Cu(I)-catalyzed azide−alkyne cycloaddition, often termed "click chemistry". We demonstrate two alkynyl-acetyl-CoA analogues, 4-pentynoyl-CoA 5-hexynoyl-CoA, function as efficient substrates lysine acetyltransferase p300 serve sensitive reagents for monitoring p300-catalyzed protein vitro. In addition, three alkynylacetate sodium 3-butynoate, 4-pentynoate, 5-hexynoate, can be metabolically incorporated onto cellular through biosynthetic mechanisms profiling cell types. Mass spectrometry analysis the enriched 4-pentynoate-labeled revealed many reported well new candidate from Jurkat T cells specific sites acetylation. The described here should powerful tools investigating