Human substance P receptor undergoes agonist‐dependent phosphorylation by G protein‐coupled receptor kinase 5 in vitro

FEBS Letters

Nishimura, Kinya³  Yamaguchi, Keisuke²  Richardson, Mark D.²  Roush, Eric D.²  Warabi, Kengo³  Barry, William T.¹  Kwatra, Madan M.² 

[1] Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, USA;Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA;Department of Anesthesiology, Juntendo University, Tokyo, Japan

关键词: Human substance P receptor;    G protein-coupled receptor kinase;    Receptor phosphorylation;    Desensitization;    GPCR;    G protein-coupled receptor;    PKA;    protein kinase A;    PKC;    protein kinase C;    GRK;    G protein-coupled receptor kinase;    Gβγ;    G protein βγ-subunit;    PLC;    phospholipase C;    hSPR;    human substance P receptor;    Sf9;    Spodoptera frugiperda;    SP;    substance P;    HEPES;    4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid;    EDTA;    ethylenediaminetetraacetic acid;    Tris–HCl;    Tris[hydroxymethyl]aminoethane hydrochloride);    PMSF;    phenylmethylsulfonyl fluoride;    SDS–PAGE;    sodium dodecyl sulfate–polyacrylamide gel electrophoresis;    AR;    adrenergic receptor;   

DOI  :  10.1016/S0014-5793(02)02858-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

G protein-coupled receptor kinases (GRKs) phosphorylate agonist-occupied G protein-coupled receptors, leading to receptor desensitization. Seven GRKs, designated GRK1 through 7, have been characterized. GRK5 is negatively regulated by protein kinase C. We investigated whether human substance P receptor (hSPR) is a substrate of GRK5. We report that membrane-bound hSPR is phosphorylated by purified GRK5, and that both the rate and extent of phosphorylation increase dramatically in the presence of substance P. The phosphorylation has a high stoichiometry (20±4 mol phosphate/mol hSPR) and a low K _m (1.7±0.1 nM). These data provide the first evidence that hSPR is a substrate of GRK5.

【 授权许可】

Unknown