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A kinetic method for measuring agonist efficacy and ligand bias using high resolution biosensors and a kinetic data analysis framework

Sci Rep. 2020; 
Hoare SRJ, Tewson PH, Quinn AM, Hughes TE.
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Peptide Synthesis Angiotensin II and SII (Sar1, Ile4,8]-Angiotensin II)40 were obtained from Genscript (Piscataway, NJ) and MyBioSource (San Diego, CA), respectively.... Trevena peptides TRV120026, TRV120045, and TRV12005547 were synthesized by Genscript. Get A Quote

摘要

The kinetics/dynamics of signaling are of increasing value for G-protein-coupled receptor therapeutic development, including spatiotemporal signaling and the kinetic context of biased agonism. Effective application of signaling kinetics to developing new therapeutics requires reliable kinetic assays and an analysis framework to extract kinetic pharmacological parameters. Here we describe a platform for measuring arrestin recruitment kinetics to GPCRs using a high quantum yield, genetically encoded fluorescent biosensor, and a data analysis framework to quantify the recruitment kinetics. The sensor enabled high temporal resolution measurement of arrestin recruitment to the angiotensin AT1 and vasopressin V2 rece... More

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