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J. Biol. Chem., Vol. 277, Issue 36, 33188-33195, September 6, 2002
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From the § Division of Digestive Diseases, Department of
Medicine, the We have previously shown that adenosine is
formed in the intestinal lumen during active inflammation from
neutrophil-derived 5'-AMP. Acting through the adenosine A2b receptor
(A2bR), the luminally derived adenosine induces vectorial chloride
secretion and a polarized secretion of interleukin-6 to the intestinal
lumen. Although some G protein-coupled receptors interact with
anchoring or signaling molecules, not much is known in this critical
area for the A2bR. We used the model intestinal epithelial cell line, T84, and Caco2-BBE cells stably transfected with GFP-A2b receptor to
study the intestinal A2bR. The A2bR is present in both the apical and
basolateral membranes of intestinal epithelia. Apical or basolateral
stimulation of the A2bR induces recruitment of the receptor to the
plasma membrane and caveolar fractions. The A2bR co-immunoprecipitates
with E3KARP and ezrin upon agonist stimulation. Ezrin interacts with
E3KARP and PKA and the interaction between ezrin and E3KARP is enhanced
by agonist stimulation. Our data suggest that the A2bR is recruited to
the plasma membrane upon apical or basolateral agonist stimulation and
interacts with E3KARP and ezrin. We speculate that such an interaction
may not only anchor the A2bR to the plasma membrane but may also
function to stabilize the receptor in a signaling complex in the plasma membrane.
The Adenosine 2b Receptor Is Recruited to the
Plasma Membrane and Associates with E3KARP and Ezrin upon Agonist
Stimulation*
§¶,,,,,, and
Epithelial Biology Unit, Department of
Pathology, and Emory University, Atlanta, Georgia 30322
*
This work was supported by National Institutes of Health
Grants K08 DK02802 (to S. V. S.), K01 DK 02831 (to D. M.), DK 35932 and DK 47662 (to J. L. M.), and the Deutsche
Forschungsgemeinschaft (to M. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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