| 26 | |
| GRIP ID | 26 |
| Main Title | Information about hetero oligomer between P2Y1 purinoceptor (P2Y1R) and A1 adenosine receptor (A1R) and interfaces of P2Y1R |
| GPCR Class | A |
| GPCR Subfamily | Purinoceptor |
| GPCR Subtype | P2Y1 |
| GPCR Name | P2Y1R |
| Partner Class | A |
| Partner Subfamily | Adenosine |
| Partner Subtype | A1 |
| Partner Name | A1R |
| Evidence | Experiment |
| Annotations | A high degree of A1R and P2Y1R colocalization was observed in cotransfected cells by double immunofluorescence experiments with confocal laser microscopy. A1R and P2Y1R coimmunoprecipitate in cotransfected HEK293T cells, suggesting that G(i/o) protein-coupled GPCRs could physically interact with Gq protein-coupled GPCRs. In the same way, A1R and P2Y2R also were found to coimmunoprecipitate in the cotransfected cells. Coexpression of A1R and P2Y1R lead to the reduction of the A1R agonist and antagonist bindings to cell membranes. A potent P2Y1R agonist adenosine 5'-O-(2-thiotriphosphate) (ADPbetaS) revealed a significant potency to A1R binding only in the cotransfected cell membranes. In addition, the A1R/P2Y1R coexpressed cells showed an ADPbetaS-dependent reduction of forskolin-evoked cAMP accumulation that was sensitive to pertussis toxin and A1R antagonist, which suggests that ADPbetaS inhibits adenylyl cyclase activity via G(i/o) proteins thorough the binding to A1R. Modified BRET observed that Myc-tagged P2Y1R fused to a Renilla luciferase (Myc-P2Y1R-Rluc) and HA-tagged A1R fused to a green fluorescent protein (HA-A1R-GFP(2)) were heterooligomerized in both constitutive and agonist-dependent manners. Addition of agonists for both A1R and P2Y1R increased the modified BRET signal in a time-dependent manner in the cells expressing HA-A1R-GFP(2) or Myc-P2Y1R-Rluc, which could be inhibited by pretreatment with the P2Y1R antagonist MRS2179. Co-localization of HA-A1R-GFP(2) and Myc-P2Y1R-Rluc was also observed in the co-transfected HEK293T cells by confocal laser microscopy. Therefore, A1R and P2Y1R are considered to form constitutive hetero-oligomers in living cells and this process is promoted by the simultaneous activation of both receptors. |
| Articles | 12123822, 11390975, 12417330 |
| Methods | Double immunofluorescence experiments, confocal laser microscopy Modified BRET |
| Target | NP_002554.1 |
| Other Partners | - |
| Disease | - |
| Interface | - |
| Mutation Information | |
| Experimentaly Information | |
| Predicted Information | |