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

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 ap...

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Bibliographic Details
Published in:FEBS letters 1986-09, Vol.206 (1), p.130-134
Main Authors: Green, Allan, Stuart, Charles A., Pietrzyk, Robert A., Partin, Maristella
Format: Article
Language:English
Subjects:
125I-HPIA, (−)-N 6-125I-p-hydroxyphenylisopropyladenosine
Adenosine
Adenosine - analogs & derivatives
Adenosine receptor
Adipocyte
Adipose Tissue - metabolism
ANB-NOS, N-5-azido-2-nitrobenzoyloxysuccinimide
Animals
Azides
Biological and medical sciences
Cell Membrane - metabolism
Cell receptors
Cell structures and functions
Cross-Linking Reagents
Fundamental and applied biological sciences. Psychology
HPIA, (−)-N 6-p-hydroxyphenylisopropyladenosine
Male
Molecular and cellular biology
Molecular Weight
NECA, 5'-(N-ethyl)carboxamidoadenosine
Phenylisopropyladenosine - analogs & derivatives
Phenylisopropyladenosine - metabolism
Photochemical cross-linking
Photochemistry
PIA, (−)-N 6-phenylisopropyladenosine
PMSF, phenylmethylsulfonyl fluoride
Rat
Rats
Rats, Inbred Strains
Receptors, Purinergic - metabolism
SANPAH, N-succinimidyl-6-(4'-azido-2'-nitrophenylamino) hexanoate
SDS-PAGE, SDS-polyacrylamide gel electrophoresis
Succinimides
sulfo-SANPAH, sulfosuccinimidyl 6-(4'-azido-2'-nitrophenylamino) hexanoate
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Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(86)81354-0