Conformational thermostabilization of the beta1-adrenergic receptor in a detergent-resistant form.

Abstract:

:There are approximately 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 beta1-adrenergic receptor. The most stable mutant receptor, betaAR-m23, contained six point mutations that led to an apparent T(m) 21 degrees C higher than the native protein, and, in the presence of bound antagonist, betaAR-m23 was as stable as bovine rhodopsin. In addition, betaAR-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.

authors

Serrano-Vega MJ,Magnani F,Shibata Y,Tate CG

doi

10.1073/pnas.0711253105

subject

Has Abstract

pub_date

2008-01-22 00:00:00

pages

877-82

issue

3

eissn

0027-8424

issn

1091-6490

pii

0711253105

journal_volume

105

pub_type

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