The Library
Engineering a minimal G protein to facilitate crystallisation of G protein-coupled receptors in their active conformation
Tools
Tools
Tools
Carpenter, Byron and Tate, Christopher G. (2016) Engineering a minimal G protein to facilitate crystallisation of G protein-coupled receptors in their active conformation. Protein Engineering Design and Selection, 29 (12). pp. 583-594. doi:10.1093/protein/gzw049
![[img]](http://wrap.warwick.ac.uk/style/images/fileicons/application_pdf.png) |
PDF
WRAP_Protein Engineering, Design and Selection-2016-Carpenter-protein-gzw049.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (1692Kb) |
Official URL: http://dx.doi.org/10.1093/protein/gzw049
Abstract
G protein-coupled receptors (GPCRs) modulate cytoplasmic signalling in response to extracellular stimuli, and are important therapeutic targets in a wide range of diseases. Structure determination of GPCRs in all activation states is important to elucidate the precise mechanism of signal transduction and to facilitate optimal drug design. However, due to their inherent instability, crystallisation of GPCRs in complex with cytoplasmic signalling proteins, such as heterotrimeric G proteins and β-arrestins, has proved challenging. Here, we describe the design of a minimal G protein, mini-Gs, which is composed solely of the GTPase domain from the adenylate cyclase stimulating G protein Gs. Mini-Gs is a small, soluble protein, which efficiently couples GPCRs in the absence of Gβγ subunits. We engineered mini-Gs, using rational design mutagenesis, to form a stable complex with detergent-solubilised β1-adrenergic receptor (β1AR). Mini G proteins induce similar pharmacological and structural changes in GPCRs as heterotrimeric G proteins, but eliminate many of the problems associated with crystallisation of these complexes, specifically their large size, conformational dynamics and instability in detergent. They are therefore novel tools, which will facilitate the biochemical and structural characterisation of GPCRs in their active conformation.
| Item Type: | Journal Article | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| Subjects: | Q Science > QP Physiology R Medicine > RS Pharmacy and materia medica |
||||||||
| Divisions: | Faculty of Science > Life Sciences (2010- ) | ||||||||
| Library of Congress Subject Headings (LCSH): | G proteins , Cell receptors, Drugs -- Design | ||||||||
| Journal or Publication Title: | Protein Engineering Design and Selection | ||||||||
| Publisher: | Oxford University Press | ||||||||
| ISSN: | 1741-0126 | ||||||||
| Official Date: | 28 December 2016 | ||||||||
| Dates: |
|
||||||||
| Date of first compliant deposit: | 12 October 2016 | ||||||||
| Volume: | 29 | ||||||||
| Number: | 12 | ||||||||
| Page Range: | pp. 583-594 | ||||||||
| DOI: | 10.1093/protein/gzw049 | ||||||||
| Status: | Peer Reviewed | ||||||||
| Publication Status: | Published | ||||||||
| Access rights to Published version: | Open Access | ||||||||
| Funder: | Heptares Therapeutics, Medical Research Council (Great Britain) (MRC) | ||||||||
| Grant number: | MRC U105197215 |
Request changes or add full text files to a record
Repository staff actions (login required)
 |
View Item |
Downloads
Downloads per month over past year
