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Molecular basis for dramatic changes in cannabinoid CB1 G protein-coupled receptor activation upon single and double point mutations

Scott, Caitlin E. and Abrol, Ravinder and Ahn, Kwang H. and Kendall, Debra A. and Goddard, William A., III (2013) Molecular basis for dramatic changes in cannabinoid CB1 G protein-coupled receptor activation upon single and double point mutations. Protein Science, 22 (1). pp. 101-113. ISSN 0961-8368. PMCID PMC3575865. https://resolver.caltech.edu/CaltechAUTHORS:20130118-083624400

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Abstract

There is considerable interest in determining the activation mechanism of G protein-coupled receptors (GPCRs), one of the most important types of proteins for intercellular signaling. Recently, it was demonstrated for the cannabinoid CB1 GPCR, that a single mutation T210A could make CB1 completely inactive whereas T210I makes it essentially constitutively active. To obtain an understanding of this dramatic dependence of activity on mutation, we used first-principles-based methods to predict the ensemble of low-energy seven-helix conformations for the wild-type (WT) and mutants (T210A and T210I). We find that the transmembrane (TM) helix packings depend markedly on these mutations, leading for T210A to both TM3+TM6 and TM2+TM6 salt-bridge couplings in the cytoplasmic face that explains the inactivity of this mutant. In contrast T210I has no such couplings across the receptor explaining the ease in activating this mutant. WT has just the TM3+TM6 coupling, known to be broken upon GPCR activation. To test this hypothesis on activity, we predicted double mutants that would convert the inactive mutant to normal activity and then confirmed this experimentally. This CB1 activation mechanism, or one similar to it, is expected to play a role in other constitutively active GPCRs as well.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1002/pro.2192DOIArticle
http://onlinelibrary.wiley.com/doi/10.1002/pro.2192/abstractPublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3575865/PubMed CentralArticle
ORCID:
AuthorORCID
Abrol, Ravinder0000-0001-7333-6793
Goddard, William A., III0000-0003-0097-5716
Additional Information:© 2012 The Protein Society. Received 5 September 2012; Accepted 26 October 2012. Published online 26 November 2012. The work in the laboratory of D.A.K. was supported by National Institutes of Health Grant DA020763. The work in the laboratory of W.A.G was initiated with gifts from PharmSelex/Accelerator and finished with funding from NIH grants (R01NS071112, R01NS073115, and R01AI040567) as well as from Sanofi.
Funders:
Funding AgencyGrant Number
NIHDA020763
NIHR01NS071112
NIHR01NS073115
NIHR01AI040567
SanofiUNSPECIFIED
Subject Keywords: protein structure prediction; G protein binding; computational methods; conformational ensemble
Issue or Number:1
PubMed Central ID:PMC3575865
Record Number:CaltechAUTHORS:20130118-083624400
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20130118-083624400
Official Citation:Scott, C. E., Abrol, R., Ahn, K. H., Kendall, D. A. and Goddard, W. A. (2013), Molecular basis for dramatic changes in cannabinoid CB1 G protein-coupled receptor activation upon single and double point mutations. Protein Science, 22: 101–113. doi: 10.1002/pro.2192
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:36463
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:18 Jan 2013 17:00
Last Modified:03 Oct 2019 04:38

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