期刊
JOURNAL OF PROTEOME RESEARCH
卷 8, 期 7, 页码 3702-3711出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr900257z
关键词
Azide; azidohomoalanine; azide-alkyne cycloaddition; cross-linking; cyclooctyne; enrichment; mass spectrometry; proteomics; peptides; pulse-chase labeling
资金
- ECHO-project
- Netherlands Organization for Scientific Research (MVO)
- National Institutes of Health [GM058867]
- National Science Foundation
- National Defense Science and Engineering
A general method is described to sequester peptides containing azides from complex peptide mixtures, aimed at facilitating mass spectrometric analysis to study different aspects of proteome dynamics. The enrichment method is based on covalent capture of azide-containing peptides by the azide-reactive cyclooctyne (ARCO) resin and is demonstrated for two different applications. Enrichment of peptides derived from cytochrome c treated with the azide-containing cross-linker bis(succinimidyl)-3-azidomethyl glutarate (BAMG) shows several cross-link containing peptides. Sequestration of peptides derived from an Escherichia coli proteome, pulse labeled with the bio-orthogonal amino acid azidohomoalanine as substitute for methionine, allows identification of numerous newly synthesized proteins. Furthermore, the method is found to be very specific, as after enrichment over 87% of all peptides contain (modified) azidohomoalanine.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据