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Expression and purification of mini G proteins from Escherichia coli
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Carpenter, Byron and Tate, Christopher G. (2017) Expression and purification of mini G proteins from Escherichia coli. Bio-protocol, 7 (8). e2235. ISSN 2331-8325.
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WRAP-expression-purification-mini-G-proteins-Carpenter-2017.pdf - Accepted Version Embargoed item. Restricted access to Repository staff only - Requires a PDF viewer. Download (1103Kb) |
Official URL: https://doi.org/10.21769/BioProtoc.2235
Abstract
Heterotrimeric G proteins modulate intracellular signalling by transducing information from cell surface G protein-coupled receptors (GPCRs) to cytoplasmic effector proteins. Structural and functional characterisation of GPCR–G protein complexes is important to fully decipher the mechanism of signal transduction. However, native G proteins are unstable and conformationally dynamic when coupled to a receptor. We therefore developed an engineered minimal G protein, mini-Gs, which formed a stable complex with GPCRs, and facilitated the crystallisation and structure determination of the human adenosine A2A receptor (A2AR) in its active conformation. Mini G proteins are potentially useful tools in a variety of applications, including characterising GPCR pharmacology, binding affinity and kinetic experiments, agonist drug discovery, and structure determination of GPCR–G protein complexes. Here, we describe a detailed protocol for the expression and purification of mini-Gs.
Item Type: | Journal Article | ||||||||||||
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Subjects: | Q Science > QD Chemistry | ||||||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||||||
Journal or Publication Title: | Bio-protocol | ||||||||||||
Publisher: | Bio-protocol LLC | ||||||||||||
ISSN: | 2331-8325 | ||||||||||||
Official Date: | 20 April 2017 | ||||||||||||
Dates: |
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Volume: | 7 | ||||||||||||
Number: | 8 | ||||||||||||
Article Number: | e2235 | ||||||||||||
Status: | Peer Reviewed | ||||||||||||
Publication Status: | Published | ||||||||||||
Copyright Holders: | Bio-protocol LLC | ||||||||||||
Date of first compliant deposit: | 25 April 2017 | ||||||||||||
Funder: | Heptares Therapeutics (Firm), Medical Research Council (Great Britain) (MRC), Wellcome Trust (London, England) | ||||||||||||
Grant number: | MRC U105197215 | ||||||||||||
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