Photochemical cross‐linking of 125I‐hydroxyphenylisopropyl adenosine to the A1 adenosine receptor of rat adipocytes

FEBS Letters

Partin, Maristella¹  Green, Allan¹  Pietrzyk, Robert A.¹  Stuart, Charles A.¹ 

[1] Division of Endocrinology, Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX 77550, USA

关键词: (Rat);    Adipocyte;    Adenosine receptor;    Adenosine;    Photochemical cross-linking;    125I-HPIA;    (−)-N 6-125I-p-hydroxyphenylisopropyladenosine;    HPIA;    (−)-N 6-p-hydroxyphenylisopropyladenosine;    PIA;    (−)-N 6-phenylisopropyladenosine;    NECA;    5'-(N-ethyl)carboxamidoadenosine;    ANB-NOS;    N-5-azido-2-nitrobenzoyloxysuccinimide;    SANPAH;    N-succinimidyl-6-(4'-azido-2'-nitrophenylamino) hexanoate;    sulfo-SANPAH;    sulfosuccinimidyl 6-(4'-azido-2'-nitrophenylamino) hexanoate;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    PMSF;    phenylmethylsulfonyl fluoride;   

DOI  :  10.1016/0014-5793(86)81354-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Rat adipocyte plasma membranes were incubated with the A₁ adenosine receptor agonist, ¹²⁵I-hydroxyphenylisopropyl adenosine (1 nM) and then treated with the photoactive cross-linking agent, ANB-NOS. The membranes were solubilized and analyzed by SDS-PAGE and autoradiography. A single protein, M _r approx. 38 000, was specifically labeled. Reduction with 2-mercaptoethanol did not affect the apparent M _r of the labeled protein. Labeling was inhibited by the adenosine receptor agonists, HPIA, PIA and NECA, and by the antagonist, theophylline, but was unaffected by inosine. We conclude that the A₁ adenosine receptor is a single protein of M _r approx. 38 000.

【 授权许可】

Unknown