Receptor selectivity between the G proteins Gα12 and Gα13 is defined by a single leucine-to-isoleucine variation

Mackenzie, A. E., Quon, T., Lin, L.-C., Hauser, A. S., Jenkins, L. , Inoue, A., Tobin, A. B. , Gloriam, D. E., Hudson, B. D. and Milligan, G. (2019) Receptor selectivity between the G proteins Gα12 and Gα13 is defined by a single leucine-to-isoleucine variation. FASEB Journal, (doi:10.1096/fj.201801956R) (PMID:30601679) (PMCID:PMC6436656) (Early Online Publication)

Mackenzie, A. E., Quon, T., Lin, L.-C., Hauser, A. S., Jenkins, L. , Inoue, A., Tobin, A. B. , Gloriam, D. E., Hudson, B. D. and Milligan, G. (2019) Receptor selectivity between the G proteins Gα12 and Gα13 is defined by a single leucine-to-isoleucine variation. FASEB Journal, (doi:10.1096/fj.201801956R) (PMID:30601679) (PMCID:PMC6436656) (Early Online Publication)

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Abstract

Despite recent advances in structural definition of GPCR–G protein complexes, the basis of receptor selectivity between G proteins remains unclear. The Gα12 and Gα13 subtypes together form the least studied group of heterotrimeric G proteins. G protein–coupled receptor 35 (GPR35) has been suggested to couple efficiently to Gα13 but weakly to Gα12. Using combinations of cells genome-edited to not express G proteins and bioluminescence resonance energy transfer–based sensors, we confirmed marked selectivity of GPR35 for Gα13. Incorporating Gα12/Gα13 chimeras and individual residue swap mutations into these sensors defined that selectivity between Gα13 and Gα12 was imbued largely by a single leucine-to-isoleucine variation at position G.H5.23. Indeed, leucine could not be substituted by other amino acids in Gα13 without almost complete loss of GPR35 coupling. The critical importance of leucine at G.H5.23 for GPR35–G protein interaction was further demonstrated by introduction of this leucine into Gαq, resulting in the gain of coupling to GPR35. These studies demonstrate that Gα13 is markedly the most effective G protein for interaction with GPR35 and that selection between Gα13 and Gα12 is dictated largely by a single conservative amino acid variation.—Mackenzie, A. E., Quon, T., Lin, L.-C., Hauser, A. S., Jenkins, L., Inoue, A., Tobin, A. B., Gloriam, D. E., Hudson, B. D., Milligan, G. Receptor selectivity between the G proteins Gα12 and Gα13 is defined by a single leucine-to-isoleucine variation.

Item Type:Articles
Status:Early Online Publication
Refereed:Yes
Glasgow Author(s) Enlighten ID:Quon, Dr Tezz and Lin, Dr Li-Chiung and Milligan, Professor Graeme and Jenkins, Mrs Laura and Hudson, Dr Brian and Tobin, Andrew and Mackenzie, Miss Amanda
Authors: Mackenzie, A. E., Quon, T., Lin, L.-C., Hauser, A. S., Jenkins, L., Inoue, A., Tobin, A. B., Gloriam, D. E., Hudson, B. D., and Milligan, G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:FASEB Journal
Publisher:Federation of American Society of Experimental Biology
ISSN:0892-6638
ISSN (Online):1530-6860
Published Online:02 January 2019
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in FASEB Journal 2019
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
726201Defining the functional roles of the enigmatic G protein coupled receptor GPR35Graeme MilliganBiotechnology and Biological Sciences Research Council (BBSRC)BB/P000649/1RI MOLECULAR CELL & SYSTEMS BIOLOGY
759351Defining the functional role of the enigmatic G protein coupled receptor GPR35 - Leicester application - PART BAndrew TobinBiotechnology and Biological Sciences Research Council (BBSRC)BB/P00069X/1RI MOLECULAR CELL & SYSTEMS BIOLOGY