Phylogeny, in situ hybridization service Sign up for PNAS Online eTocs
Link: Info for AuthorsLink: Editorial BoardLink: AboutLink: SubscribeLink: AdvertiseLink: ContactLink: Sitemap Link: PNAS Home
Proceedings of the National Academy of Sciences
Link: Current Issue "" Link: Archives "" Link: Online Submission ""  Link: Advanced Search

Published online on January 11, 2008, 10.1073/pnas.0711253105
PNAS | January 22, 2008 | vol. 105 | no. 3 | 877-882


This Article
Right arrow Figures Only
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supporting Information
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a colleague
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My File Cabinet
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via CrossRef
Google Scholar
Right arrow Articles by Serrano-Vega, M. J.
Right arrow Articles by Tate, C. G.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Serrano-Vega, M. J.
Right arrow Articles by Tate, C. G.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg  
What's this?

 Previous Article  | Table of Contents |  Next Article 

BIOLOGICAL SCIENCES / BIOCHEMISTRY
Conformational thermostabilization of the β1-adrenergic receptor in a detergent-resistant form

Maria J. Serrano-Vega, Francesca Magnani, Yoko Shibata, and Christopher G. Tate*

Laboratory of Molecular Biology, Medical Research Council, Hills Road, Cambridge CB2 0QH, United Kingdom

Communicated by Richard Henderson, Medical Research Council, Cambridge, United Kingdom, November 28, 2007 (received for review November 7, 2007)

There are {approx}350 non-odorant G protein-coupled receptors (GPCRs) encoded by the human genome, many of which are predicted to be potential therapeutic targets, but there are only two structures available to represent the whole of the family. We hypothesized that improving the detergent stability of these receptors and simultaneously locking them into one preferred conformation will greatly improve the chances of crystallization. We developed a generic strategy for the isolation of detergent-solubilized thermostable mutants of a GPCR, the β1-adrenergic receptor. The most stable mutant receptor, βAR-m23, contained six point mutations that led to an apparent Tm 21°C higher than the native protein, and, in the presence of bound antagonist, βAR-m23 was as stable as bovine rhodopsin. In addition, βAR-m23 was significantly more stable in a wide range of detergents ideal for crystallization and was preferentially in an antagonist conformation in the absence of ligand.

G protein-coupled receptor | membrane protein | stabilization

Author contributions: M.J.S.-V. and C.G.T. designed research; M.J.S.-V. performed all of the experiments; F.M. and Y.S. contributed significantly to method development and the concept of conformational thermostabilization as a generic methodology through their own research on other GPCRs; M.J.S.-V. analyzed data; and C.G.T. wrote the paper.

The authors declare no conflict of interest.

This article contains supporting information online at www.pnas.org/cgi/content/full/0711253105/DC1.

*To whom correspondence should be addressed. E-mail: cgt{at}mrc-lmb.cam.ac.uk

© 2008 by The National Academy of Sciences of the USA


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg    What's this?