定量重建膜蛋白为脂膜蛋白体的原理。

Rationale for the Quantitative Reconstitution of Membrane Proteins into Proteoliposomes.

机构信息

Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, CNRS UMR 7099, Institut de Biologie Physico-Chimique (IBPC), Université Paris Diderot, Sorbonne Paris Cité, PSL Research University, Paris, France.

CBMN UMR-CNRS 5248 University of Bordeaux, Pessac, France.

出版信息

Methods Mol Biol. 2020;2168:63-72. doi: 10.1007/978-1-0716-0724-4_3.

DOI:10.1007/978-1-0716-0724-4_3

PMID:33582987

Abstract

Proteoliposome reconstitution is a method of choice for the investigation of membrane proteins as it allows their manipulation in the desired hydrophobic environment and allows one to tackle their study from both functional and structural points of view. Methods for their rapid and efficient reconstitution have been known for a long time but the quality and dispersity of the resulting suspensions is often overlooked. Here we describe our routine for the obtention of monodisperse populations of proteoliposomes as well as for the quantitation of protein per liposome.

摘要

脂质体重建是研究膜蛋白的首选方法，因为它允许在所需的疏水环境中对其进行操作，并允许从功能和结构两个方面来研究它们。长期以来，人们已经知道了快速有效地重建它们的方法，但人们往往忽略了所得到悬浮液的质量和分散性。在这里，我们描述了我们获得单分散脂质体群体以及定量脂质体中蛋白质的常规程序。