Rapid and improved reconstitution of bacterial mechanosensitive ion channel proteins MscS and MscL into liposomes using a modified sucrose method

被引：51

作者：

Battle, Andrew R. ^{[1
]}

Petrov, Evgeny ^{[1
]}

Pal, Prithwish ^{[1
]}

Martinac, Boris ^{[1
]}

机构：

[1] Univ Queensland, Sch Biomed Sci, Dept Physiol & Pharmacol, Brisbane, Qld 4072, Australia

来源：

FEBS LETTERS | 2009年 / 583卷 / 02期

基金：

澳大利亚研究理事会; 英国医学研究理事会;

关键词：

Patch clamp; Confocal microscopy; Fluorescence; Lipid; Sugar; ESCHERICHIA-COLI; MEMBRANE-PROTEINS; GIANT LIPOSOMES; SINGLE RESIDUE; PURIFICATION; STABILIZATION; MECHANISM; PRESSURE;

D O I：

10.1016/j.febslet.2008.12.033

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：

The bacterial mechanosensitive (MS) channels of small (MscS) and large (MscL) conductance have functionally been reconstituted into giant unilamellar liposomes (GUVs) using an improved reconstitution method in the presence of sucrose. This method gives significant time savings (preparation times as little as 6 h) compared to the classical method of protein reconstitution which uses a dehydration/rehydration (D/R) procedure (minimum 2 days preparation time). Moreover, it represents the first highly reproducible method for functional reconstitution of MscS as well as MscS/MscL co-reconstitution. This novel procedure has the potential to be used for studies of other ion channels by liposome reconstitution. (C) 2008 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.

引用

页码：407 / 412

页数：6

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