Mass spectrometry of membrane transporters reveals subunit stoichiometry and interactions

被引：143

作者：

Barrera, Nelson P. ^{[1
]}

Isaacson, Shoshanna C. ^{[1
]}

Zhou, Min ^{[1
]}

Bavro, Vassiliy N. ^{[2
]}

Welch, Alex ^{[3
]}

Schaedler, Theresia A. ^{[3
]}

Seeger, Markus A. ^{[3
]}

Miguel, Ricardo Nunez ^{[4
]}

Korkhov, Vladimir M. ^{[5
]}

van Veen, Hendrik W. ^{[3
]}

Venter, Henrietta ^{[3
]}

Walmsley, Adrian R. ^{[6
]}

Tate, Christopher G. ^{[5
]}

Robinson, Carol V. ^{[1
]}

机构：

[1] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England

[2] Univ Oxford, Dept Phys, Clarendon Lab, Oxford OX1 3PU, England

[3] Univ Cambridge, Dept Pharmacol, Cambridge CB2 1QJ, England

[4] Univ Cambridge, Dept Biochem, Cambridge CB2 1QW, England

[5] MRC, Mol Biol Lab, Cambridge CB2 2QH, England

[6] Univ Durham, Sch Biol & Biomed Sci, Durham, England

来源：

NATURE METHODS | 2009年 / 6卷 / 08期

基金：

英国医学研究理事会; 英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

MULTIDRUG TRANSPORTER; ESCHERICHIA-COLI; LACTOCOCCUS-LACTIS; ABC TRANSPORTER; PSEUDOMONAS-AERUGINOSA; EMRE; ASSEMBLIES; EXPRESSION; COMPLEXES; MECHANISM;

D O I：

10.1038/nmeth.1347

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a general mass spectrometry approach to determine subunit stoichiometry and lipid binding in intact membrane protein complexes. By exploring conditions for preserving interactions during transmission into the gas phase and for optimally stripping away detergent, by subjecting the complex to multiple collisions, we released the intact complex largely devoid of detergent. This enabled us to characterize both subunit stoichiometry and lipid binding in 4 membrane protein complexes.

引用

页码：585 / U49

页数：5

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