Direct analysis of protein complexes using mass spectrometry

被引:1838
|
作者
Link, AJ
Eng, J
Schieltz, DM
Carmack, E
Mize, GJ
Morris, DR
Garvik, BM
Yates, JR [1 ]
机构
[1] Univ Washington, Dept Mol Biotechnol, Seattle, WA 98195 USA
[2] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
[3] Fred Hutchinson Canc Res Inst, Seattle, WA 98109 USA
关键词
protein identification; mass spectrometry; multidimensional chromatography; ribosome; yeast genome;
D O I
10.1038/10890
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We describe a rapid, sensitive process for comprehensively identifying proteins in macromolecular complexes that uses multidimensional liquid chromatography (LC) and tandem mass spectrometry (MS/MS) to separate and fragment peptides. The SEQUEST algorithm, relying upon translated genomic sequences, infers amino acid sequences from the fragment ions. The method was applied to the Saccharomyces cerevisiae ribosome leading to the identification of a novel protein component of the yeast and human 40S subunit. By offering the ability to identify >100 proteins in a single run, this process enables components in even the largest macromolecular complexes to be analyzed comprehensively.
引用
收藏
页码:676 / 682
页数:7
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