测量NLR寡聚化III：通过生物发光共振能量转移检测NLRP3复合物

Measuring NLR Oligomerization III: Detection of NLRP3 Complex by Bioluminescence Resonance Energy Transfer.

作者信息

Martín-Sánchez Fátima, Compan Vincent, Pelegrín Pablo

机构信息

Inflammation and Experimental Surgery Unit,, Institute for Bio-Health Research of Murcia (IMIB-Arrixaca), Carretera Buenavista, s/n., El Palmar, Murcia, 30120, Spain.

Institut de Génomique Fonctionnelle, Labex ICST, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Université Montpellier, Montpellier, France.

出版信息

Methods Mol Biol. 2016;1417:159-68. doi: 10.1007/978-1-4939-3566-6_10.

DOI:10.1007/978-1-4939-3566-6_10

PMID:27221488

Abstract

Bioluminescent resonance energy transfer (BRET) is a natural phenomenon resulting from a non-radiative energy transfer between a bioluminescent donor (Renilla luciferase) and a fluorescent protein acceptor. BRET signal is dependent on the distance and the orientation between the donor and the acceptor and could be used to study protein-protein interactions and conformational changes within proteins in real time in living cells. This protocol describes the use of BRET technique to study NLRP3 oligomerization in living cells before and during NLRP3 inflammasome activation.

摘要

生物发光共振能量转移（BRET）是一种自然现象，由生物发光供体（海肾荧光素酶）与荧光蛋白受体之间的非辐射能量转移产生。BRET信号取决于供体与受体之间的距离和方向，可用于实时研究活细胞内蛋白质-蛋白质相互作用以及蛋白质的构象变化。本方案描述了使用BRET技术研究NLRP3炎性小体激活之前和激活过程中活细胞内NLRP3的寡聚化。