biostudies-literatureChen HNIAID NIH HHSNHLBI NIH HHSNIGMS NIH HHSeabn0067https://www.ebi.ac.uk/biostudies/studies/S-EPMC8967229biostudies-literatureUnknown8(13)Sphingosine-1-phosphate (S1P) regulates immune cell trafficking, angiogenesis, and vascular function via its five receptors. Inherited mutations in S1P receptor 2 (S1PR2) occur in individuals with hearing loss, and acquired mutations in S1PR2 and G_α13 occur in a malignant lymphoma. Here, we present the cryo-electron microscopy structure of S1P-bound S1PR2 coupled to the heterotrimeric G₁₃. Interaction between S1PR2 intracellular loop 2 (ICL2) and transmembrane helix 4 confines ICL2 to engage the α5 helix of G_α13. Transforming growth factor-α shedding assays and cell migration assays support the key roles of the residues in S1PR2-G_α13 complex assembly. The structure illuminates the mechanism of receptor disruption by disease-associated mutations. Unexpectedly, we showed that FTY720-P, an agonist of the other four S1PRs, can trigger G₁₃ activation via S1PR2. S1PR2^F274I variant can increase the activity of G₁₃ considerably with FTY720-P and S1P, thus revealing a basis for S1PR drug selectivity.Science advancesStructure of S1PR2-heterotrimeric G₁₃ signaling complex.PMC8967229R01 AI045073P01 HL020948R01 GM135343Cyster JGLi XChen KHuang WChen HStaudt LMfalseStructure of S1PR2-heterotrimeric G₁₃ signaling complex.Sphingosine-1-phosphate (S1P) regulates immune cell trafficking, angiogenesis, and vascular function via its five receptors. Inherited mutations in S1P receptor 2 (S1PR2) occur in individuals with hearing loss, and acquired mutations in S1PR2 and G_α13 occur in a malignant lymphoma. Here, we present the cryo-electron microscopy structure of S1P-bound S1PR2 coupled to the heterotrimeric G₁₃. Interaction between S1PR2 intracellular loop 2 (ICL2) and transmembrane helix 4 confines ICL2 to engage the α5 helix of G_α13. Transforming growth factor-α shedding assays and cell migration assays support the key roles of the residues in S1PR2-G_α13 complex assembly. The structure illuminates the mechanism of receptor disruption by disease-associated mutations. Unexpectedly, we showed that FTY720-P, an agonist of the other four S1PRs, can trigger G₁₃ activation via S1PR2. S1PR2^F274I variant can increase the activity of G₁₃ considerably with FTY720-P and S1P, thus revealing a basis for S1PR drug selectivity.2022-01-01T00:00:00Z2022 Apr2026-05-10T01:36:56.992Z2025-02-19T01:10:43.238ZS-EPMC89672293535355910.1126/sciadv.abn0067