Role of G protein-coupled receptor kinases (GRKs) in β 2 -adrenoceptor-mediated glucose uptake.

Autor: Ham S; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Mukaida S; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Sato M; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Keov P; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Bengtsson T; Atrogi AB, Stockholm, Sweden.; Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden., Furness S; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Holliday ND; School of Life Sciences, The Medical School, Queen's Medical Centre, University of Nottingham, Nottingham, UK.; Excellerate Bioscience, Biocity, Nottingham, UK., Evans BA; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Summers RJ; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia., Hutchinson DS; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
Jazyk: angličtina
Zdroj: Pharmacology research & perspectives [Pharmacol Res Perspect] 2024 Feb; Vol. 12 (1), pp. e1176.
DOI: 10.1002/prp2.1176
Abstrakt: Truncation of the C-terminal tail of the β 2 -AR, transfection of βARKct or over-expression of a kinase-dead GRK mutant reduces isoprenaline-stimulated glucose uptake, indicating that GRK is important for this response. We explored whether phosphorylation of the β 2 -AR by GRK2 has a role in glucose uptake or if this response is related to the role of GRK2 as a scaffolding protein. CHO-GLUT4myc cells expressing wild-type and mutant β 2 -ARs were generated and receptor affinity for [ 3 H]-CGP12177A and density of binding sites determined together with the affinity of isoprenaline and BRL37344. Following receptor activation by β 2 -AR agonists, cAMP accumulation, GLUT4 translocation, [ 3 H]-2-deoxyglucose uptake, and β 2 -AR internalization were measured. Bioluminescence resonance energy transfer was used to investigate interactions between β 2 -AR and β-arrestin2 or between β 2 -AR and GRK2. Glucose uptake after siRNA knockdown or GRK inhibitors was measured in response to β 2 -AR agonists. BRL37344 was a poor partial agonist for cAMP generation but displayed similar potency and efficacy to isoprenaline for glucose uptake and GLUT4 translocation. These responses to β 2 -AR agonists occurred in CHO-GLUT4myc cells expressing β 2 -ARs lacking GRK or GRK/PKA phosphorylation sites as well as in cells expressing the wild-type β 2 -AR. However, β 2 -ARs lacking phosphorylation sites failed to recruit β-arrestin2 and did not internalize. GRK2 knock-down or GRK2 inhibitors decreased isoprenaline-stimulated glucose uptake in rat L6 skeletal muscle cells. Thus, GRK phosphorylation of the β 2 -AR is not associated with isoprenaline- or BRL37344-stimulated glucose uptake. However, GRKs acting as scaffold proteins are important for glucose uptake as GRK2 knock-down or GRK2 inhibition reduces isoprenaline-stimulated glucose uptake.
(© 2024 The Authors. Pharmacology Research & Perspectives published by British Pharmacological Society and American Society for Pharmacology and Experimental Therapeutics and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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