Dynamics of apelin receptor/G protein coupling in living cells |
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Authors: | Bo Bai Yunlu Jiang Xin Cai Jing Chen |
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Affiliation: | 1. Neurobiology Institute, Jining Medical University, Jining 272067, PR China;2. Division of Metabolic and Vascular Health, Warwick Medical School, University of Warwick, Coventry, UK |
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Abstract: | During our research on apelin receptor (APJ) signalling in living cells with BRET and FRET, we demonstrated that apelin-13 stimulation can lead to the activation of Gαi2 or Gαi3 through undergoing a molecular rearrangement rather than dissociation in HEK293 cells expressing APJ. Furthermore, Gαo and Gαq also showed involvement in APJ activation through a classical dissociation model. However, both FRET signal and BRET ratio between fluorescent Gαi1 subunit and Gβγ subunits demonstrated little change after apelin-13 stimulation. These results demonstrated that stimulation of APJ with apelin-13 causes activation of Gαi2, Gαi3, Gαo, Gαq; among which Gαi2, Gαi3 were activated through a novel rearrangement process. These results provide helpful data for understanding APJ mediated G-protein signalling. |
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Keywords: | GPCR, G protein-coupled receptor APJ, apelin receptor HEK, human embryonic kidney BRET, bioluminescence resonance energy transfer FRET, Fö rster resonance energy transfer Rluc, Renilla reniformis luciferase YFP, yellow fluorescent protein CFP, cyan fluorescent protein ELISA, Enzyme-Linked Immunosorbent Assay |
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