G-protein-coupled A1 adenosine receptors in coated vesicles of mammalian brain: characterization by radioligand binding and photoaffinity labelling.

  title={G-protein-coupled A1 adenosine receptors in coated vesicles of mammalian brain: characterization by radioligand binding and photoaffinity labelling.},
  author={G Gonz{\'a}lez-Calero and Ana Cubero and Karl-Norbert Klotz},
  journal={Cellular signalling},
  volume={4 6},
A1 adenosine receptors in coated vesicles have been characterized by radioligand binding and photoaffinity labelling. Saturation experiments with the antagonist 8-cyclopentyl-1,3-[3H]dipropyl-xanthine ([3H]DPCPX) gave a Kd value of 0.7 nM and a Bmax value of 82 +/- 13 fmol/mg protein. For the highly A1-selective agonist 2-chloro-N6-[3H]cyclopentyladenosine ([3H]CCPA) a Kd value of 1.7 nM and a Bmax value of 72 +/- 29 fmol/mg protein was estimated. Competition of agonists for [3H]DPCPX binding… Expand
6 Citations
Binding of an adenosine A1 receptor agonist and adenosine A1 receptor antagonist to sheep pineal membranes.
2-chloro-cyclopentyl-[3H]-adenosine and [3H]DPCPX bind specifically to sheep pineal membranes to elucidate the significance of the differences observed between the binding properties of the adenosine A1 receptor agonist and adenosines A1 receptors. Expand
Desensitization and internalization of adenosine A1 receptors in rat brain by in vivo treatment with R-PIA: involvement of coated vesicles.
Results demonstrate adenosine A1 receptor desensitization in rat brain by 'in vivo' treatment with R-PIA and suggest a role for coated vesicles in the internalization of G-protein coupled receptors. Expand
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Bovine Brain Coated Vesicles Contain Adenosine A1 Receptors. Presence of Adenylate Cyclase Coupled to the Receptor
The results of this study support the evidence that adenosine A1 receptors present in coated vesicles are coupled to adenylate cyclase activity through a Gi protein. Expand
Co-purification of A1 adenosine receptors and guanine nucleotide-binding proteins from bovine brain.
Following removal of 8-p-sulfophenyltheophylline and GTP, purified receptors bind agonists and antagonists with affinities similar to crude soluble receptors and typical of A1 receptors. Expand
Coupling of adenosine A1 receptors to a G-protein in coated vesicles isolated from bovine brain: presence of pertussis and cholera toxin substrates.
Results confirm the presence of GTP binding proteins (alpha Gi and alpha Gs) in coated vesicles isolated from bovine brain. Expand
Photoaffinity labeling of A1-adenosine receptors.
The results indicate that 125I-AHPIA identifies the ligand-binding subunit of the A1-adenosine receptor, which is a peptide with Mr = 35,000, which can be blocked specifically and stereoselectively by adenosine receptors agonists and antagonists in a manner which is typical for the A 1-subtype. Expand
Characterization of the Solubilized A1 Adenosine Receptor from Rat Brain Membranes
The results suggest that the solubilized A1 adenosine receptors are still coupled to the guanine nucleotide binding protein Ni and that all regulatory functions are retained on Solubilization. Expand
Evidence for the presence of muscarinic acetylcholine receptors in bovine brain coated vesicles.
Evidence obtained from quinuclidinylbenzilate binding determinations suggested that muscarinic acetylcholine receptor molecules are present in purified bovine brain coated vesicles, and binding competition experiments using atropine, N-methylscopolamine, oxotremorine, and carbamyl choline confirmed the typicalMuscarinic nature of the binding site. Expand
Detection and characterization of beta-adrenergic receptors and adenylate cyclase in coated vesicles isolated from bovine brain
The results suggest that receptors present in coated vesicles are cryptic in nature, and beta-adrenergic receptor binding to peaks I and II was measured with 125I-cyanopindolol (CYP) as ligand in Sepharose 4B column assays. Expand
The glycoprotein nature of A1 adenosine receptors.
  • K. Klotz, M. Lohse
  • Chemistry, Medicine
  • Biochemical and biophysical research communications
  • 1986
A1 adenosine receptors from different tissues and species were photoaffinity labelled and the carbohydrate content was examined by both enzymatic and chemical treatment showing that neuraminidase treatment alters the electrophoretic mobility of the receptor band indicating the presence of terminal neuraminic acids. Expand
Stereospecific opiate-binding sites occur in coated vesicles
P purified bovine brain-coated vesicles contained high affinity stereospecific opiate alkaloid-binding sites with characteristic opioid binding properties. Expand
Solubilized adenosine receptors in the brain: regulation of guanine nucleotides.
The apparent retention of a complex of receptors and guanosine triphosphate binding but not cation binding protein in the soluble state may permit a molecular analysis of receptor regulation. Expand