Proteomics on full-length membrane proteins using mass spectrometry

被引：82

作者：

le Coutre, J

Whitelegge, JP

Gross, A

Turk, E

Wright, EM

Kaback, HR

Faull, KF

机构：

[1] Univ Calif Los Angeles, Howard Hughes Med Inst, Dept Phys, MacDonald Res Labs 6720, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, Howard Hughes Med Inst, Dept Microbiol & Mol Genet, Los Angeles, CA 90095 USA

[3] Univ Calif Los Angeles, Inst Mol Biol, Los Angeles, CA 90095 USA

[4] Univ Calif Los Angeles, Dept Biochem & Chem, Los Angeles, CA 90095 USA

[5] Univ Calif Los Angeles, Pasarow Mass Spectrometry Lab, Los Angeles, CA 90095 USA

[6] Univ Calif Los Angeles, Jules Stein Eye Inst, Los Angeles, CA 90095 USA

[7] Univ Calif Los Angeles, Dept Physiol, Los Angeles, CA 90095 USA

[8] Univ Calif Los Angeles, Dept Psychiat & Biobehav Sci, Los Angeles, CA 90095 USA

[9] Univ Calif Los Angeles, Inst Neuropsychiat, Los Angeles, CA 90095 USA

来源：

BIOCHEMISTRY | 2000年 / 39卷 / 15期

关键词：

D O I：

10.1021/bi000150m

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A general technique has been developed that allows rapid mass spectrometric analysis of full-length membrane proteins [Whitelegge, J. P., le Coutre, J., et al. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 10695-10698], Using in-line HPLC electrospray ionization mass spectrometry (LC-MS), different native and recombinant bacterial membrane proteins of up to 61 kDa are characterized. Mass spectrometric data of four entirely different membrane proteins from three bacterial organisms, two transporters, a channel, and a porin protein are presented. In addition to determination of the molecular mass with an accuracy of +/-0.01%, the technique monitors alkylation or oxidation of single Cys residues and errors in deduced amino acid sequences. Finally, using in-line LC-MS, unknown proteins can be identified from solubilized Escherichia coli membranes without prior purification.

引用

页码：4237 / 4242

页数：6

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