Quantitation of de novo localized 15N-labeled lipoproteins and membrane proteins having one and two transmembrane segments in a Bacillus subtilis secA temperature-sensitive mutant using 2D-PAGE and MALDI-TOF MS

被引：9

作者：

Bunai, K

Nozaki, M

Kakeshita, H

Nemoto, T

Yamane, K

机构：

[1] RIKEN, Discovery Res Inst, Cellular Dynam Lab, Wako, Saitama 3510198, Japan

[2] Univ Tsukuba, Inst Biol Sci, Tsukuba, Ibaraki 3058572, Japan

[3] Natl Inst Adv Ind Sci & Technol, Tsukuba, Ibaraki 3058566, Japan

[4] Natl Food Res Inst, Tsukuba, Ibaraki 3058642, Japan

来源：

JOURNAL OF PROTEOME RESEARCH | 2005年 / 4卷 / 03期

关键词：

proteomics; membrane protein localization; SecA; stable isotope N-15; Bacillus subtilis; MALDI-TOF MS; 2D-PAGE;

D O I：

10.1021/pr049755l

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We developed a means of quantifying proteins that have just localized in the cytoplasmic membrane using N-15-whole cell labeling together with 2D-PAGE and MALDI-TOF MS. The localization of 18 among 20 proteins consisting of 8 lipoproteins, 11 integral membrane proteins having one or two transmembrane segments and one secretory protein in the membrane fractions of Bacillus subtilis, was inhibited by the absence of SecA in a temperature-sensitive mutant. The time course of inhibition indicated that SecA participates in the localization of those proteins through immediately dependent, delayed dependent, and independent ways.

引用

页码：826 / 836

页数：11