Close association of B2 bradykinin receptors with P2Y2 ATP receptors

Two G-protein-coupled receptors (GPCRs) that couple with G alpha q/11, B2 bradykinin (BK) receptor (B2R) and ATP/UTP receptor P2Y(2) (P2Y(2)R), are ubiquitously expressed and responsible for vascular tone, inflammation, and pain. We analysed the cellular signalling of P2Y(2)Rs in cells that express B2Rs. B2R desensitization induced by BK or B2R internalization-inducing glycans cross-desensitized the P2Y(2)R response to ATP/UTP. Fluorescence resonance energy transfer from P2Y(2)R-AcGFP to B2R-DsRed was detected in the cells and on the cell surfaces, showing the close association of these GPCRs. BK- and ATP-induced cross-internalization of P2Y(2)R and B2R, respectively, was shown in a beta-galactosidase complementation assay using P2Y(2)R or B2R fused to the H31R substituted a donor peptide of a beta-galactosidase reporter enzyme (P2Y(2)R-alpha or B2R-alpha) with coexpression of the FYVE domain of endofin, an early endosome protein, fused to the M15 acceptor deletion mutant of p-galactosidase (the omega peptide, FYVE-omega). Arrestin recruitment to the GPCRs by cross-activation was also shown with the similar way. Coimmunoprecipitation showed that B2R and P2Y(2)R were closely associated in the cotransfected cells. These results indicate that B2R couples with P2Y(2)R and that these GPCRs act together to fine-tune cellular responsiveness. The collaboration between these receptors may permit rapid onset and turning off of biological events.