Sequential Peptide Affinity (SPA) system for the identification of mammalian and bacterial protein complexes

被引：157

作者：

Zeghouf, M

Li, J

Butland, G

Borkowska, A

Canadien, V

Richards, D

Beattie, B

Emili, A

Greenblatt, JF

机构：

[1] Univ Toronto, Banting & Best Dept Med Res, Toronto, ON M5G 1L6, Canada

[2] Univ Toronto, Dept Mol & Med Genet, Toronto, ON M5S 1A8, Canada

[3] Affinium Pharmaceut, Toronto, ON M5J 1V6, Canada

来源：

JOURNAL OF PROTEOME RESEARCH | 2004年 / 3卷 / 03期

关键词：

SPA system; protein complexes; protein-protein interactions; affinity purification; mass spectrometry; human cell; E; coli;

D O I：

10.1021/pr034084x

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A vector system is described that combines reliable, very low level, regulated protein expression in human cells with two affinity purification tags (Sequential Peptide Affinity, or SPA, system). By avoiding overproduction of the target protein, this system allows for the efficient purification of natural protein complexes and their identification by mass spectrometry. We also present an adaptation of the SPA system for the efficient purification and identification of protein complexes in E. coli and, potentially, other bacteria.

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页码：463 / 468

页数：6

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