| Literature DB >> 26749445 |
Pawel K Dominik1, Marta T Borowska1, Olivier Dalmas1, Sangwoo S Kim1, Eduardo Perozo1, Robert J Keenan1, Anthony A Kossiakoff2.
Abstract
A major challenge in membrane biophysics is to define the mechanistic linkages between a protein's conformational transitions and its function. We describe a novel approach to stabilize transient functional states of membrane proteins in native-like lipid environments allowing for their structural and biochemical characterization. This is accomplished by combining the power of antibody Fab-based phage display selection with the benefits of embedding membrane protein targets in lipid-filled nanodiscs. In addition to providing a stabilizing lipid environment, nanodiscs afford significant technical advantages over detergent-based formats. This enables the production of a rich pool of high-performance Fab binders that can be used as crystallization chaperones, as fiducial markers for single-particle cryoelectron microscopy, and as probes of different conformational states. Moreover, nanodisc-generated Fabs can be used to identify detergents that best mimic native membrane environments for use in biophysical studies.Entities:
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Year: 2015 PMID： 26749445 PMCID： PMC4740257 DOI： 10.1016/j.str.2015.11.014
Source DB: PubMed Journal: Structure ISSN： 0969-2126 Impact factor: 5.006