The Library
Strategy for the thermostabilization of an agonist-bound GPCR coupled to a G protein
Tools
Strege, Annette, Carpenter, Byron, Edwards, Patricia C. and Tate, Christopher G. (2017) Strategy for the thermostabilization of an agonist-bound GPCR coupled to a G protein. In: Ziegler, Christine, (ed.) Methods In Enzymology. A Structure-Function Toolbox for Membrane Transporter and Channels, 594 . Elsevier, pp. 243-264. ISBN 9780128123539 (In Press)
Research output not available from this repository.
Request-a-Copy directly from author or use local Library Get it For Me service.
Official URL: https://doi.org/10.1016/bs.mie.2017.05.014
Abstract
Structure determination of G protein-coupled receptors (GPCRs) in the inactive state bound to high-affinity antagonists has been very successful through the implementation of a number of protein engineering and crystallization strategies. However, the structure determination of GPCRs in their fully active state coupled to a G protein is still very challenging. Recently, mini-G proteins were developed, which recapitulate the coupling of a full heterotrimeric G protein to a GPCR despite being less than one-third of the size. This allowed the structure determination of the agonist-bound adenosine A2A receptor (A2AR) coupled to mini-Gs. Although this is extremely encouraging, A2AR is very stable compared with many other GPCRs, particularly when an agonist is bound. In contrast, the agonist-bound conformation of the human corticotropin-releasing factor receptor is considerably less stable, impeding the formation of good quality crystals for structure determination. We have therefore developed a novel strategy for the thermostabilization of a GPCR–mini-G protein complex. In this chapter, we will describe the theoretical and practical principles of the thermostability assay for stabilizing this complex, discuss its strengths and weaknesses, and show some typical results from the thermostabilization process.
Item Type: | Book Item | ||||
---|---|---|---|---|---|
Subjects: | Q Science > QD Chemistry | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Series Name: | A Structure-Function Toolbox for Membrane Transporter and Channels | ||||
Journal or Publication Title: | Methods in Enzymology | ||||
Publisher: | Elsevier | ||||
ISBN: | 9780128123539 | ||||
ISSN: | 978-0-12-812353-9 | ||||
Book Title: | Methods In Enzymology | ||||
Editor: | Ziegler, Christine | ||||
Official Date: | 19 July 2017 | ||||
Dates: |
|
||||
Volume: | 594 | ||||
Page Range: | pp. 243-264 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | In Press | ||||
Copyright Holders: | Elsevier | ||||
Funder: | Medical Research Council (Great Britain) (MRC), Heptares Therapeutics (Firm) | ||||
Grant number: | MC_U105197215 |
Request changes or add full text files to a record
Repository staff actions (login required)
View Item |