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Immunological identification of A 1 adenosine receptors in brain cortex
Author(s) -
Ciruela F.,
Casadó V.,
Mallol J.,
Canela E. I.,
Lluis C.,
Franco R.
Publication year - 1995
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490420610
Subject(s) - adenosine , receptor , immunoprecipitation , adenosine receptor , extracellular , antiserum , adenosine a2b receptor , intracellular , microbiology and biotechnology , ligand (biochemistry) , biochemistry , biology , adenosine a1 receptor , chemistry , antibody , immunology , agonist , gene
The A 1 adenosine receptor from pig brain cortex has been identified by means of two antipeptide antibodies against two domains of the receptor molecule: PC/10 antiserum was raised against a part of the third intracellular loop, and PC/20 antiserum was raised against a part of the second extracellular loop. PC/10 antibody was able to recognize a 39‐kDa band that corresponded to the A l receptor, as demonstrated by immunoblotting and by immunoprecipitation of the molecule cross‐linked to [ 125 I](R)‐2‐azidoN 2 ‐p‐hydroxy(phenylisopropyl)adenosine. Besides the 39‐kDa band, PC/20 also recognized a 74‐kDa form that does not seem to correspond to a receptor‐G protein complex. The occurrence of the two bands was detected and analyzed in samples from different species and tissues showing a heterogeneous distribution of both. The 74‐kDa form can be converted into the 39‐kDa form by treatment with agonists or antagonists of A l adenosine receptors. These results suggest that A 1 adenosine receptor can occur in dimers and that the dimer–monomer conversion might be regulated by adenosine as the physiological ligand. Since the 74‐kDa aggregates were not recognized by PC/10, it is likely that part of the third intracellular loop participates in the protein–protein interaction. ©1995 Wiley‐Liss, Inc.