Combining Metabolic Alkyne Labeling and Click Chemistry for Secretome Analysis of Serum-Containing C

• 作 者: Jiangnan Zheng Yuan Mao Shun Feng Ruijun Tian
• 机 构: Department of Chemistry,School of Science,Southern University of Science and Technology,Shenzhen,Gua
• 出 处: 中国化学（英文版）
• 发表日期: 2021年7期
• 关键词: Click chemistry Mass spectrometry Proteomics Secretome Metabolic labeling

Main observation and conclusionrnBioorthogonal click chemistry has emerged as a powerful tool for the specific modification of proteins in complex mixtures.Metabolic labeling of proteins with azide followed by the copper-catalyzed azide-alkyne cycloaddition (CuAAC) with alkyne-based affinity probes/beads is widely applied to study protein turnover and post-translational modifications (PTMs).However,it has long been known that the alkyne-based enrichment of high concentration protein samples (e.g.,serum) is not sufficient to remove high abun-dant contaminating proteins.Herein,we demonstrate that the protein contamination is mainly caused by thiol-yne addition between terminal alkyne and cysteine residue.Furthermore,we report that azide-based enrichment combined with metabolic labeling with alkyne can significantly reduce contamination and improve the enrichment efficiency of secreted proteins in serum-containing condi-tioned medium.