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Volume 270, Number 28, Issue of July 14, pp. 16895-16902, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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Differential Interaction with and Regulation of Multiple G-proteins by the Rat A Adenosine Receptor

Timothy M. Palmer , Thomas W. Gettys , Gary L. Stiles

Interaction of the rat A adenosine receptor (AAR) with G-proteins has been assessed using a stably transfected Chinese hamster ovary cell system. The non-selective AR agonist 5`-N-ethylcarboxamidoadenosine (NECA) increased the labeling of a 41-kDa membrane protein by 4-azidoanilido-[-P]guanosine 5`-triphosphate (AA-[P]GTP), a photolabile GTP analogue. Subsequent immunoprecipitation of G -subunits indicated that NECA stimulated incorporation of label into both G-2 and G-3. Additional experiments revealed an AAR stimulation of label into G and/or G -subunits, albeit to a lesser degree than that elicited by endogenous P purinergic receptors. No interaction with G could be detected. Sustained cellular exposure to NECA induced AAR desensitization and specific down-regulation of G-3 and G-protein -subunits without changing levels of G-2, G, or G -subunits. Therefore the AAR can interact with G-2, G-3, and, to some extent, G-like proteins, but sustained agonist exposure down-regulates only one of the G-proteins with which it interacts. This is the first description of the differing specificities of AAR/G-protein coupling versus down-regulation in situ and provides a potential mechanism by which the AAR could elicit the heterologous desensitization of signaling events mediated by G3.

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