A Simple Method for the Reconstitution of Membrane Proteins into Giant Unilamellar Vesicles

被引：0
作者：
Armelle Varnier
Frédérique Kermarrec
Iulia Blesneac
Christophe Moreau
Lavinia Liguori
Jean Luc Lenormand
Nathalie Picollet-D’hahan
机构：
[1] CEA,HumProTher Laboratory
[2] DSV,undefined
[3] iRTSV,undefined
[4] Biopuces,undefined
[5] IBS (CEA,undefined
[6] CNRS,undefined
[7] UJF),undefined
[8] Protéines Membranaires,undefined
[9] TheREx,undefined
[10] TIMC-IMAG,undefined
[11] CNRS UMR5525,undefined
[12] UJF,undefined
[13] UFR de Médecine,undefined
来源：
Journal of Membrane Biology | 2010年 / 233卷
关键词：
Biomembranes; Membrane protein; Membrane channel; Giant unilamellar vesicle; Proteoliposome; Fusion; Lipid;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
A simple method for the reconstitution of membrane protein from submicron proteoliposomes into giant unilamellar vesicles (GUVs) is presented here: This method does not require detergents, fusion peptides or a dehydration step of the membrane protein solution. In a first step, GUVs of lipids were formed by electroformation, purified and concentrated; and in a second step, the concentrated GUV solution was added to a small volume of vesicles or proteoliposomes. Material transfer from submicron vesicles and proteoliposomes to GUVs occurred spontaneously and was characterized with fluorescent microscopy and patch-clamp recordings. As a functional test, the voltage-dependent, anion-selective channel protein was reconstituted into GUVs, and its electrophysiological activity was monitored with the patch clamp. This method is versatile since it is independent of the presence of the protein, as demonstrated by the fusion of fluorescently labeled submicron vesicles and proteoliposomes with GUVs.
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页码：85 / 92
页数：7
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