Proteomics on full-length membrane proteins using mass spectrometry
Proteomics on full-length membrane proteins using mass spectrometry
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DOI：
10.1021/bi000150m
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发表时间：
2000-04-18
期刊：
影响因子：
2.9
通讯作者：
Faull, KF
中科院分区：
文献类型：
--
作者：
le Coutre, J;Whitelegge, JP;Faull, KF
已经开发了一种通用技术，其允许对全长膜蛋白进行快速质谱分析[Whitelegge，J. P.，le Coutre，J.，等人（1999）Proc.Natl. Acad. Sci. U.S.A.96，10695-10698]，使用在线HPLC电喷雾电离质谱（LC-MS），表征了高达61 kDa的不同天然和重组细菌膜蛋白。质谱数据的四个完全不同的膜蛋白从三个细菌生物体，两个转运蛋白，通道，和孔蛋白。除了以+/-0.01%的准确度测定分子量外，该技术还监测单个Cys残基的烷基化或氧化以及推导的氨基酸序列中的错误。最后，使用在线LC-MS，未知蛋白质可以从溶解的大肠杆菌膜中鉴定，而无需事先纯化。
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.