A novel complete reconstitution system for membrane integration of the simplest membrane protein
A novel complete reconstitution system for membrane integration of the simplest membrane protein
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DOI：
10.1016/j.bbrc.2010.03.061
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发表时间：
2010-04-09
影响因子：
3.1
通讯作者：
Tokuda, Hajime
中科院分区：
文献类型：
--
作者：
Nishiyama, Ken-ichi;Maeda, Masahide;Tokuda, Hajime
通过确定的因子开发了最简单蛋白质膜整合的完整重构系统 Pf3 外壳蛋白的突变版本 3L-Pf3 外壳不需要信号识别颗粒/Sec 因子，也不需要膜电位即可整合到大肠杆菌的细胞质膜中 虽然 3L-Pf3 外壳自发整合到由磷脂组成的脂质体中，但二酰甘油在生理水平上完全阻断这种自发整合 在自发整合不发生的条件下如果没有发生，3L-Pf3 涂层整合绝对依赖于一种新的整合刺激因子。 PURE系统是由纯化的大肠杆菌中参与翻译的因子组成的体外翻译系统，与含有高度纯化的整合刺激因子的脂质体相结合，揭示了3L-Pf3外壳整合的多个循环，实现了膜整合的完全重构。根据该因子的功能，我们建议将该因子命名为MPlase（膜蛋白整合酶）。 (C) 2010 爱思唯尔公司保留所有权利
A complete reconstitution system for membrane integration of the simplest protein was developed by means of defined factors A mutant version of Pf3 coat protein, 3L-Pf3 coat, requires neither signal recognition particle/Sec factors nor a membrane potential for its integration into the cytoplasmic membrane of Escherichia colt Although 3L-Pf3 coat is spontaneously integrated into liposomes composed of phospholipids, diacylglycerol completely blocks such spontaneous integrations at a physiological level Under the conditions where spontaneous integration does not occur, 3L-Pf3 coat integration was absolutely dependent on a novel integration-stimulating factor. Combination of the PURE system, an in vitro translation system composed of the purified factors involved in translation in E. coli, with liposomes containing the highly purified integration-stimulating factor revealed multiple cycles of 3L-Pf3 coat integration, achieving the complete reconstitution of membrane integration Based on the function of the factor, we propose that the factor is named MPlase (Membrane Protein Integrase). (C) 2010 Elsevier Inc All rights reserved