Membrane protein reconstitution in nanodiscs for luminescence spectroscopy studies

Nanotechnology Reviews

article

Maria E. Zoghbi¹  Guillermo A. Altenberg² 

[1] School of Natural Sciences, University of California, 4225 N. Hospital Road;Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center

关键词: ATP-binding cassette;    LRET;    luminescence resonance energy transfer;    MsbA;    multidrug resistance;   

DOI  :  10.1515/ntrev-2016-0078

学科分类：社会科学、人文和艺术（综合）

来源: De Gruyter

【 摘 要 】

ATP-binding cassette (ABC) exporters transport substrates across biological membranes using ATP hydrolysis by a process that involves switching between inward- and outward-facing conformations. Most of the structural studies of ABC proteins have been performed with proteins in detergent micelles, locked in specific conformations and/or at low temperature. In this article, we present recent data from our laboratories where we studied the prototypical ABC exporter MsbA during ATP hydrolysis, at 37°C, reconstituted in a lipid bilayer. These studies were possible through the use of luminescence resonance energy transfer spectroscopy in MsbA reconstituted in nanodiscs. We found major differences between MsbA in these native-like conditions and in previous studies. These include a separation between the nucleotide-binding domains that was much smaller than previously thought, and a large fraction of molecules with associated nucleotide-binding domains in the nucleotide-free apo state. These studies stress the importance of studying membrane proteins in an environment that approaches physiological conditions.

【 授权许可】

CC BY