![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
ME Olah, C Gallo-Rodriguez, KA Jacobson and GL Stiles
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710.
The rat A3 adenosine receptor (AR) is a recently characterized AR subtype cloned from testis and brain cDNA libraries. N6-2-(4-Amino-3- [125I]iodophenyl)ethyladenosine, a high affinity A1AR agonist, has served as the only radioligand available for study of the A3AR. The relatively low affinity of N6-2-(4-amino-3-[125I] iodophenyl)ethyladenosine for the A3AR and its greater A1AR selectivity necessitate the development of more appropriate radioligands for A3AR analysis. This report characterizes 125I-4-aminobenzyl-5'-N- methylcarboxamidoadenosine (125I-AB-MECA), a high affinity radioligand for the A3AR, in two cell lines that express this AR subtype. Membranes from Chinese hamster ovary (CHO) cells expressing the rat A3AR and from the rat mast cell line RBL-2H3 bound 125I-AB-MECA with Kd values of 1.48 +/- 0.33 nM and 3.61 +/- 0.30 nM, respectively. As determined by 125I-AB-MECA binding, levels of A3AR expression in the A3AR-CHO cell line and RBL-2H3 cells were 3.06 +/- 0.21 pmol/mg and 1.02 +/- 0.13 pmol/mg, respectively. Binding of 125I-AB-MECA was characterized in competition assays. In the A3AR-CHO cell line a potency order of cyclohexyl-5'-N-ethylcarboxamidoadenosine (cyclohexyl-NECA) = benzyl- NECA > (-)-N6-[(R)-phenylisopropyl]adenosine = NECA was observed, and in RBL-2H3 cells (-)-N6-[(R)-phenylisopropyl]adenosine and NECA were equipotent. Xanthine amine congener (XAC) and 8-cyclopentyl-1,3- dipropylxanthine did not significantly inhibit 125I-AB-MECA binding. The parent compound, AB-MECA, dose-dependently inhibited forskolin- stimulated adenylyl cyclase activity in A3AR-CHO cell membranes. 125I- AB-MECA bound to the rat A1AR and canine A2aAR expressed in COS-7 cells with Kd values of 3.42 +/- 0.43 nM and 25.1 +/- 12.6 nM, respectively. This binding was significantly reduced in the presence of 1 microM XAC. In RBL-2H3 cells, XAC had no effect on 125I-AB-MECA affinity and reduced the level of radioligand binding by approximately 5%.
This article has been cited by other articles:
|
|
 |
|
  D. van der Hoeven, T. C. Wan, and J. A. Auchampach Activation of the A3 Adenosine Receptor Suppresses Superoxide Production and Chemotaxis of Mouse Bone Marrow Neutrophils Mol. Pharmacol., September 1, 2008; 74(3): 685 - 696. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z.-D. Ge, J. N. Peart, L. M. Kreckler, T. C. Wan, M. A. Jacobson, G. J. Gross, and J. A. Auchampach Cl-IB-MECA [2-Chloro-N6-(3-iodobenzyl)adenosine-5'-N-methylcarboxamide] Reduces Ischemia/Reperfusion Injury in Mice by Activating the A Adenosine Receptor J. Pharmacol. Exp. Ther., December 1, 2006; 319(3): 1200 - 1210. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Ernens, D. Rouy, E. Velot, Y. Devaux, and D. R. Wagner Adenosine Inhibits Matrix Metalloproteinase-9 Secretion By Neutrophils: Implication of A2a Receptor and cAMP/PKA/Ca2+ Pathway Circ. Res., September 15, 2006; 99(6): 590 - 597. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Jajoo, D. Mukherjea, S. Pingle, Y. Sekino, and V. Ramkumar Induction of Adenosine A1 Receptor Expression by Pertussis Toxin via an Adenosine 5'-Diphosphate Ribosylation-Independent Pathway J. Pharmacol. Exp. Ther., April 1, 2006; 317(1): 1 - 10. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Kreckler, T. C. Wan, Z.-D. Ge, and J. A. Auchampach Adenosine Inhibits Tumor Necrosis Factor-{alpha} Release from Mouse Peritoneal Macrophages via A2A and A2B but Not the A3 Adenosine Receptor J. Pharmacol. Exp. Ther., April 1, 2006; 317(1): 172 - 180. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. C. Pingle, S. Mishra, A. Marcuzzi, S. G. Bhat, Y. Sekino, L. P. Rybak, and V. Ramkumar Osmotic Diuretics Induce Adenosine A1 Receptor Expression and Protect Renal Proximal Tubular Epithelial Cells against Cisplatin-mediated Apoptosis J. Biol. Chem., October 8, 2004; 279(41): 43157 - 43167. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.M. Weiss, K. Benwell, I.A. Cliffe, R.J. Gillespie, A.R. Knight, J. Lerpiniere, A. Misra, R.M. Pratt, D. Revell, R. Upton, et al. Discovery of nonxanthine adenosine A2A receptor antagonists for the treatment of Parkinson's disease Neurology, December 9, 2003; 61(90116): S101 - 106. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. P. Headrick, B. Hack, and K. J. Ashton Acute adenosinergic cardioprotection in ischemic-reperfused hearts Am J Physiol Heart Circ Physiol, November 1, 2003; 285(5): H1797 - H1818. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Trincavelli, D. Tuscano, M. Marroni, A. Falleni, V. Gremigni, S. Ceruti, M. P. Abbracchio, K. A. Jacobson, F. Cattabeni, and C. Martini A3 Adenosine Receptors in Human Astrocytoma Cells: Agonist-Mediated Desensitization, Internalization, and Down-Regulation Mol. Pharmacol., December 1, 2002; 62(6): 1373 - 1384. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. T. Warren, P. D. Andrews, C. W. Gourlay, and K. R. Ayscough Sla1p couples the yeast endocytic machinery to proteins regulating actin dynamics J. Cell Sci., April 15, 2002; 115(8): 1703 - 1715. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. B. Fredholm, A. P. IJzerman, K. A. Jacobson, K.-N. Klotz, and J. Linden International Union of Pharmacology. XXV. Nomenclature and Classification of Adenosine Receptors Pharmacol. Rev., December 1, 2001; 53(4): 527 - 552. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-G. Gao, J. E. Van Muijlwijk-Koezen, A. Chen, C. E. Muller, A. P. Ijzerman, and K. A. Jacobson Allosteric Modulation of A3 Adenosine Receptors by a Series of 3-(2-Pyridinyl)isoquinoline Derivatives Mol. Pharmacol., November 1, 2001; 60(5): 1057 - 1063. [Abstract] [Full Text]
|
|
|
|
 |
|
 S. J. Mundell, M. E. Olah, R. A. Panettieri Jr., J. L. Benovic, and R. B. Penn Regulation of G Protein-Coupled Receptor-Adenylyl Cyclase Responsiveness in Human Airway Smooth Muscle by Exogenous and Autocrine Adenosine Am. J. Respir. Cell Mol. Biol., February 1, 2001; 24(2): 155 - 163. [Abstract] [Full Text]
|
|
|
|
|
|
K. Varani, S. Merighi, S. Gessi, K.-N. Klotz, E. Leung, P. G. Baraldi, B. Cacciari, R. Romagnoli, G. Spalluto, and P. A. Borea [3H]MRE 3008F20: A Novel Antagonist Radioligand for the Pharmacological and Biochemical Characterization of Human A3 Adenosine Receptors Mol. Pharmacol., May 1, 2000; 57(5): 968 - 975. [Abstract] [Full Text]
|
|
|
|
|
|
T. M. Palmer and G. L. Stiles Identification of Threonine Residues Controlling the Agonist-Dependent Phosphorylation and Desensitization of the Rat A3 Adenosine Receptor Mol. Pharmacol., March 1, 2000; 57(3): 539 - 545. [Abstract] [Full Text]
|
|
|
|
|
|
F Santini, R. Penn, A. Gagnon, J. Benovic, and J. Keen Selective recruitment of arrestin-3 to clathrin coated pits upon stimulation of G protein-coupled receptors J. Cell Sci., January 7, 2000; 113(13): 2463 - 2470. [Abstract] [PDF]
|
|
|
|
|
|
R. D. Lasley, P. Narayan, M. S. Jahania, E. L. Partin, K. R. Kraft, and R. M. Mentzer Jr. Species-dependent hemodynamic effects of adenosine A3-receptor agonists IB-MECA and Cl-IB-MECA Am J Physiol Heart Circ Physiol, June 1, 1999; 276(6): H2076 - H2084. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Ralevic and G. Burnstock Receptors for Purines and Pyrimidines Pharmacol. Rev., September 1, 1998; 50(3): 413 - 492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Auchampach, X. Jin, T. C. Wan, G. H. Caughey, and J. Linden Canine Mast Cell Adenosine Receptors: Cloning and Expression of the A3 Receptor and Evidence that Degranulation Is Mediated by the A2B Receptor Mol. Pharmacol., November 1, 1997; 52(5): 846 - 860. [Abstract] [Full Text]
|
|
|
|
|
|
T. M. Palmer, C. A. Harris, J. Coote, and G. L. Stiles Induction of Multiple Effects on Adenylyl Cyclase Regulation by Chronic Activation of the Human A3 Adenosine Receptor Mol. Pharmacol., October 1, 1997; 52(4): 632 - 640. [Abstract] [Full Text]
|
|
|
|
|
|
M. E. Olah and M. E. Olah Identification of A2a Adenosine Receptor Domains Involved in Selective Coupling to GS. ANALYSIS OF CHIMERIC A1/A2a ADENOSINE RECEPTORS J. Biol. Chem., January 3, 1997; 272(1): 337 - 344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Palmer, J. L. Benovic, and G. L. Stiles Molecular Basis for Subtype-specific Desensitization of Inhibitory Adenosine Receptors. ANALYSIS OF A CHIMERIC A1-A3 ADENOSINE RECEPTOR J. Biol. Chem., June 21, 1996; 271(25): 15272 - 15278. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Palmer, J. L. Benovic, and G. L. Stiles Agonist-dependent Phosphorylation and Desensitization of the Rat A(3) Adenosine Receptor J. Biol. Chem., December 8, 1995; 270(49): 29607 - 29613. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Palmer, T. W. Gettys, and G. L. Stiles Differential Interaction with and Regulation of Multiple G-proteins by the Rat A[IMAGE] Adenosine Receptor J. Biol. Chem., July 14, 1995; 270(28): 16895 - 16902. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Jacobson, R. Xie, L. Young, L. Chang, and B. T. Liang A Novel Pharmacological Approach to Treating Cardiac Ischemia. BINARY CONJUGATES OF A1 AND A3 ADENOSINE RECEPTOR AGONISTS J. Biol. Chem., September 22, 2000; 275(39): 30272 - 30279. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
