{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Tatsumi M"],"funding":["Takeda Science Foundation","Japan Agency for Medical Research and Development","National Institutes of Health","Japan Science and Technology Agency","NIGMS NIH HHS","NIH HHS","Japan Society for the Promotion of Science"],"pagination":["11119"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11096383"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["14(1)"],"pubmed_abstract":["G-protein-coupled receptors (GPCRs) transduce diverse signals into the cell by coupling to one or several Gα subtypes. Of the 16 Gα subtypes in human cells, Gα<sub>12</sub> and Gα<sub>13</sub> belong to the G<sub>12</sub> subfamily and are reported to be functionally different. Notably, certain GPCRs display selective coupling to either Gα<sub>12</sub> or Gα<sub>13</sub>, highlighting their significance in various cellular contexts. However, the structural basis underlying this selectivity remains unclear. Here, using a Gα<sub>12</sub>-coupled designer receptor exclusively activated by designer drugs (DREADD; G<sub>12</sub>D) as a model system, we identified residues in the α5 helix and the receptor that collaboratively determine Gα<sub>12</sub>-vs-Gα<sub>13</sub> selectivity. Residue-swapping experiments showed that G<sub>12</sub>D distinguishes differences between Gα<sub>12</sub> and Gα<sub>13</sub> in the positions G.H5.09 and G.H5.23 in the α5 helix. Molecular dynamics simulations observed that I378<sup>G.H5.23</sup> in Gα<sub>12</sub> interacts with N103<sup>2.39</sup>, S169<sup>3.53</sup> and Y176<sup>34.53</sup> in G<sub>12</sub>D, while H364<sup>G.H5.09</sup> in Gα<sub>12</sub> interact with Q264<sup>5.71</sup> in G<sub>12</sub>D. Screening of mutations at these positions in G<sub>12</sub>D identified G<sub>12</sub>D mutants that enhanced coupling with Gα<sub>12</sub> and to an even greater extent with Gα<sub>13</sub>. Combined mutations, most notably the dual Y176<sup>34.53</sup>H and Q264<sup>5.71</sup>R mutant, further enhanced Gα<sub>12</sub><sub>/</sub><sub>13</sub> coupling, thereby serving as a potential Gα<sub>12/13</sub>-DREADD. Such novel Gα<sub>12/13</sub>-DREADD may be useful in future efforts to develop drugs that target Gα<sub>12/13</sub> signaling as well as to identify their therapeutic indications."],"journal":["Scientific reports"],"pubmed_title":["Identification of Gα<sub>12</sub>-vs-Gα<sub>13</sub>-coupling determinants and development of a Gα<sub>12/13</sub>-coupled designer GPCR."],"pmcid":["PMC11096383"],"funding_grant_id":["JP22J10475","SC2GM130480","P21H04791, JP21H05113, JPJSBP120213501 and JPJSBP120218801","JPMJFR215T, JPMJMS2023 and 22714181","SC2 GM130480","JP22ama121038 and JP22zf0127007"],"pubmed_authors":["Ikuta T","Cruz C","Inoue A","Tatsumi M","Nakamura G","Kamakura N","Abrol R","Kuwabara R"],"additional_accession":[]},"is_claimable":false,"name":"Identification of Gα<sub>12</sub>-vs-Gα<sub>13</sub>-coupling determinants and development of a Gα<sub>12/13</sub>-coupled designer GPCR.","description":"G-protein-coupled receptors (GPCRs) transduce diverse signals into the cell by coupling to one or several Gα subtypes. Of the 16 Gα subtypes in human cells, Gα<sub>12</sub> and Gα<sub>13</sub> belong to the G<sub>12</sub> subfamily and are reported to be functionally different. Notably, certain GPCRs display selective coupling to either Gα<sub>12</sub> or Gα<sub>13</sub>, highlighting their significance in various cellular contexts. However, the structural basis underlying this selectivity remains unclear. Here, using a Gα<sub>12</sub>-coupled designer receptor exclusively activated by designer drugs (DREADD; G<sub>12</sub>D) as a model system, we identified residues in the α5 helix and the receptor that collaboratively determine Gα<sub>12</sub>-vs-Gα<sub>13</sub> selectivity. Residue-swapping experiments showed that G<sub>12</sub>D distinguishes differences between Gα<sub>12</sub> and Gα<sub>13</sub> in the positions G.H5.09 and G.H5.23 in the α5 helix. Molecular dynamics simulations observed that I378<sup>G.H5.23</sup> in Gα<sub>12</sub> interacts with N103<sup>2.39</sup>, S169<sup>3.53</sup> and Y176<sup>34.53</sup> in G<sub>12</sub>D, while H364<sup>G.H5.09</sup> in Gα<sub>12</sub> interact with Q264<sup>5.71</sup> in G<sub>12</sub>D. Screening of mutations at these positions in G<sub>12</sub>D identified G<sub>12</sub>D mutants that enhanced coupling with Gα<sub>12</sub> and to an even greater extent with Gα<sub>13</sub>. Combined mutations, most notably the dual Y176<sup>34.53</sup>H and Q264<sup>5.71</sup>R mutant, further enhanced Gα<sub>12</sub><sub>/</sub><sub>13</sub> coupling, thereby serving as a potential Gα<sub>12/13</sub>-DREADD. Such novel Gα<sub>12/13</sub>-DREADD may be useful in future efforts to develop drugs that target Gα<sub>12/13</sub> signaling as well as to identify their therapeutic indications.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 May","modification":"2026-06-01T11:58:36.424Z","creation":"2026-04-08T12:09:22.658Z"},"accession":"S-EPMC11096383","cross_references":{"pubmed":["38750247"],"doi":["10.1038/s41598-024-61506-4"]}}