期刊
FASEB JOURNAL
卷 33, 期 4, 页码 5005-5017出版社
FEDERATION AMER SOC EXP BIOL
DOI: 10.1096/fj.201801956R
关键词
GPCR; GPR35; G protein barcode; genome editing
资金
- Novo Nordisk Foundation [NNF17OC0031226]
- European Research Council [639125]
- U.K. Medical Research Council
- Glasgow University Leadership Fellowship
- U.K. Biotechnology and Biosciences Research Council [BB/P000649/1, BB/P00069X/1]
- Japanese Society for the Promotion of Science [17K08264]
- Japan Agency for Medical Research and Development Grant [PRIME JP17gm5910013]
- BBSRC [BB/P00069X/1, BB/P000649/1] Funding Source: UKRI
- MRC [MR/P019366/1, MC_UP_A600_1110] Funding Source: UKRI
- European Research Council (ERC) [639125] Funding Source: European Research Council (ERC)
Despite recent advances in structural definition of GPCR-G protein complexes, the basis of receptor selectivity between G proteins remains unclear. The G alpha(12) and G alpha(13) subtypes together form the least studied group of heterotrimeric G proteins. G protein-coupled receptor 35 (GPR35) has been suggested to couple efficiently to G alpha(13) but weakly to G alpha(12). 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 G alpha(13). Incorporating G alpha(12)/G alpha(13) chimeras and individual residue swap mutations into these sensors defined that selectivity between G alpha(13) and G alpha(12) 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 G alpha(13) 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 G alpha(q), resulting in the gain of coupling to GPR35. These studies demonstrate that G alpha(13) is markedly the most effective G protein for interaction with GPR35 and that selection between G alpha(13) and G alpha(12) is dictated largely by a single conservative amino acid variation.
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