Publication Date:
2015-07-29
Description:
Two G-protein-coupled receptors (GPCRs) that couple with Gαq/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 β-galactosidase complementation assay using P2Y 2 R or B2R fused to the H31R substituted α donor peptide of a β-galactosidase reporter enzyme (P2Y 2 R-α or B2R-α) with coexpression of the FYVE domain of endofin, an early endosome protein, fused to the M15 acceptor deletion mutant of β-galactosidase (the peptide, FYVE-). 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.
Print ISSN:
0021-924X
Electronic ISSN:
1756-2651
Topics:
Biology
,
Chemistry and Pharmacology
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