Selective Enrichment of Azide-Containing Peptides from Complex Mixtures
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- 作者：Merel A. Nessen ; Gertjan Kramer ; JaapWillem Back ; Jeremy M. Baskin ; Linde E. J. Smeenk ; Leo J. de Koning ; Jan H. van Maarseveen ; Luitzen de Jong ; Carolyn R. Bertozzi ; Henk Hiemstra ; Chris G. de Koster
- 刊名：Journal of Proteome Research
- 出版年：2009
- 出版时间：July 6, 2009
- 年：2009
- 卷：8
- 期：7
- 页码：3702-3711
- 全文大小：226K
- 年卷期：v.8,no.7(July 6, 2009)
- ISSN：1535-3907
文摘
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.