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Fingerprinting of GPCR activity across multiple G protein substrates unveils complex profiles of functional selectivity (1066.12)
Author(s) -
Masuho Ikuo,
Jones Christopher,
Martemyanov Kirill
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1066.12
Subject(s) - g protein coupled receptor , receptor , g protein , drug discovery , signal transduction , functional selectivity , computational biology , chemistry , biology , microbiology and biotechnology , biochemistry
Despite the tractability of G protein‐coupled receptors (GPCRs) as drug targets, there are substantial challenges in understanding the mechanisms of drug action at these receptors. Since more than 800 GPCRs and 16 Gα subunits are identified in human genome and are capable to generate enormous array of GPCR‐G protein combinations, G protein‐coupling profiles of GPCRs are a critical missing piece of the GPCR‐signaling puzzle. Here, we have undertaken an attempt to comprehensively characterize G protein‐coupling selectivity and its modulation by agonists and intracellular regulatory proteins. We employed a single‐platform BRET assay to evaluate GPCR responses via 14 different Gα subunits using dynamic association/dissociation of Venus‐tagged Gβγ with NanoLuc‐tagged GRK3‐derived sensor in living cells. Distinct G protein‐coupling profiles for different receptors in both maximal amplitudes and kinetics of responses were observed and used as a basis for modeling “fingerprints” for each of the examined receptors. These fingerprints in many cases differed between physiological agonists and synthetic ligands. We further found that members of Regulators of G protein Signaling (RGS) family shape GPCR fingerprints. These results reveal underappreciated complexity of GPCR coupling to different G protein substrates and suggest a new avenue for designing signaling pathway‐specific therapeutics. Grant Funding Source : Supported by the NIH grants HL105550 and NS081282.