The implications of proteolytic background for shotgun proteomics

被引:148
作者
Picotti, Paola
Aebersold, Ruedi
Domon, Bruno
机构
[1] ETH, Inst Mol Syst Biol, CH-8093 Zurich, Switzerland
[2] Univ Zurich, Fac Sci, CH-8006 Zurich, Switzerland
[3] Inst Mol Syst Biol, Seattle, WA 98103 USA
关键词
D O I
10.1074/mcp.M700029-MCP200
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The analysis by liquid chromatography coupled to tandem mass spectrometry of complex peptide mixtures, generated by proteolysis of protein samples, is the main proteomics method used today. The approach is based on the assumption that each protein present in a sample reproducibly and predictably generates a relatively small number of peptides that can be identified by mass spectrometry. In this study this assumption was examined by a targeted peptide sequencing strategy using inclusion lists to trigger peptide fragmentation attempts. It was found that the number of peptides observed from a single protein is at least one order of magnitude greater than previously assumed. This unexpected complexity of proteomics samples implies substantial technical challenges, explains some perplexing results in the proteomics literature, and prompts the need for developing alternative experimental strategies for the rapid and comprehensive analysis of proteomes.
引用
收藏
页码:1589 / 1598
页数:10
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