Human substance P receptor expressed in Chinese hamster ovary cells directly activates Gαq/11, Gαs, and Gαo

FEBS Letters

Kwatra, Madan M¹  Roush, Eric D¹ 

[1] Departments of Anesthesiology and Pharmacology, P.O. Box 3094, Duke University Medical Center, Durham, NC 27710, USA

关键词: Human substance P receptor;    G protein;    Chinese hamster ovary cell;    [32P]AA-GTP;    P3-(4-azidoanilido)-[P1-32P]-5′-guanosine triphosphate;    cAMP;    cyclic adenosine 3′:5′-cyclic monophosphate;    CHO;    Chinese hamster ovary;    CHO-hSPR;    Chinese hamster ovary cell stably transfected with hSPR;    CHO-K1;    Chinese hamster ovary cells;    strain K1;    SDS-PAGE;    sodium dodecylsulfate-polyacrylamide gel electrophoresis;    SP;    substance P;    SPR;    substance P receptor;    hSPR;    human substance P receptor;   

DOI  :  10.1016/S0014-5793(98)00553-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Substance P receptor (SPR) stably expressed in Chinese hamster ovary (CHO) cells stimulates at least three second messenger systems including phosphoinositide hydrolysis, cyclic AMP (cAMP) formation, and arachidonic acid release. Whether these second messenger systems are activated via single or multiple G proteins is not known. Therefore, in the present study we examined whether human SPR (hSPR) stably expressed in CHO cells activates multiple G proteins. This was achieved by photoaffinity labeling of G_α-subunits with [³²P]azidoanilido-GTP ([³²P]AA-GTP) upon hSPR stimulation in CHO-hSPR membranes followed by immunoprecipitation of the labeled G_α-subunits with antibodies specific for various G_α-subunits. These experiments reveal that hSPR directly activates G_αq/11, G_αs and G_αo. While hSPR is known to couple G_αq/11, the present study provides the first evidence that hSPR can also activate G_αs and G_αo in a mammalian system.

【 授权许可】

Unknown