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