Endocrinology, Vol 135, 2130-2136, Copyright © 1994 by Endocrine Society

ARTICLES

Short-term desensitization of the angiotensin II receptor of bovine adrenal glomerulosa cells corresponds to a shift from a high to a low affinity state

G Boulay, L Chretien, DE Richard and G Guillemette
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.

Angiotensin II (Ang II) is an important regulator of aldosterone production by bovine adrenal glomerulosa (BAG) cells. Ang II interacts with a specific receptor coupled to a guanyl nucleotide-binding protein (G protein) that controls the activity of phospholipase C. A primary culture of BAG cells was used to study short-term desensitization of the Ang II receptor. After short exposures to Ang II, BAG cells lost some [125I]Ang II binding capacity. This loss was dependent on the duration of the pretreatment and on the concentration of Ang II used. A maximal loss of [125I]Ang II binding of 55 +/- 10% was observed after a pretreatment of 30 min with 30 nM Ang II. The EC50 was 1.3 +/- 0.6 nM (mean +/- SD of three experiments). The desensitization was readily reversible, since most of the binding capacity (higher than 90%) was recovered after a 60-min incubation, at 37 C, in the absence of Ang II. Scatchard studies revealed that the Ang II receptor of BAG cells exists under two affinity states with one dissociation constant of 0.2 nM and another dissociation constant of 1.5 nM. After a 30-min exposure of BAG cells to 10 nM Ang II, an important decrease of high affinity binding sites was observed. The maximal amount of binding sites was similar on control and desensitized cells (around 52,000 receptors per cell). GTP gamma S, a potent activator of G proteins, decreased [125I]Ang II binding to permeabilized BAG cells. This GTP gamma S effect was not observed on permeabilized BAG cells that had previously been desensitized with 10 nM Ang II. These results suggested that, similarly to GTP gamma S, short exposure to 10 nM Ang II caused the uncoupling of Ang II receptor from its G protein. DuP-753 (a selective AT1 angiotensin II type 1 receptor antagonist) markedly unhibited, whereas PD-123319 (a selective AT2 angioten II type 2 receptor antagonist) had no effect on Ang II receptor desensitization, indicating that the AT1 receptor subtype was responsible for the observed phenomenon. Pretreatment of BAG cells with staurosporine (a protein kinase C inhibitor) and R24571 (a calmodulin inhibitor) did not modify Ang II- induced desensitization of AT1 receptor.(ABSTRACT TRUNCATED AT 400 WORDS)

This article has been cited by other articles:

				D. G Romero, M. Y. Zhou, L. L Yanes, M. W Plonczynski, T. R Washington, C. E Gomez-Sanchez, and E. P Gomez-Sanchez Regulators of G-protein signaling 4 in adrenal gland: localization, regulation, and role in aldosterone secretion J. Endocrinol., August 1, 2007; 194(2): 429 - 440. [Abstract] [Full Text] [PDF]

				J. D. Violin, S. M. DeWire, W. G. Barnes, and R. J. Lefkowitz G Protein-coupled Receptor Kinase and beta-Arrestin-mediated Desensitization of the Angiotensin II Type 1A Receptor Elucidated by Diacylglycerol Dynamics J. Biol. Chem., November 24, 2006; 281(47): 36411 - 36419. [Abstract] [Full Text] [PDF]

				M. Auger-Messier, G. Arguin, B. Chaloux, R. Leduc, E. Escher, and G. Guillemette Down-Regulation of Inositol 1,4,5-Trisphosphate Receptor in Cells Stably Expressing the Constitutively Active Angiotensin II N111G-AT1 Receptor Mol. Endocrinol., December 1, 2004; 18(12): 2967 - 2980. [Abstract] [Full Text] [PDF]

				A. SPAT and L. HUNYADY Control of Aldosterone Secretion: A Model for Convergence in Cellular Signaling Pathways Physiol Rev, April 1, 2004; 84(2): 489 - 539. [Abstract] [Full Text] [PDF]

				M. Auger-Messier, M. Clement, P. M. Lanctot, P. C. Leclerc, R. Leduc, E. Escher, and G. Guillemette The Constitutively Active N111G-AT1 Receptor for Angiotensin II Maintains a High Affinity Conformation Despite Being Uncoupled from Its Cognate G Protein Gq/11{alpha} Endocrinology, December 1, 2003; 144(12): 5277 - 5284. [Abstract] [Full Text] [PDF]

				P. C. Leclerc, M. Auger-Messier, P. M. Lanctot, E. Escher, R. Leduc, and G. Guillemette A Polyaromatic Caveolin-Binding-Like Motif in the Cytoplasmic Tail of the Type 1 Receptor for Angiotensin II Plays an Important Role in Receptor Trafficking and Signaling Endocrinology, December 1, 2002; 143(12): 4702 - 4710. [Abstract] [Full Text] [PDF]

				S. Miserey-Lenkei, C. Parnot, S. Bardin, P. Corvol, and E. Clauser Constitutive Internalization of Constitutively Active Angiotensin II AT1A Receptor Mutants Is Blocked by Inverse Agonists J. Biol. Chem., February 15, 2002; 277(8): 5891 - 5901. [Abstract] [Full Text] [PDF]

				J. J. Saris, M. M.E.D. van den Eijnden, J. M.J. Lamers, P. R. Saxena, M. A.D.H. Schalekamp, and A.H. J. Danser Prorenin-Induced Myocyte Proliferation: No Role for Intracellular Angiotensin II Hypertension, February 1, 2002; 39(2): 573 - 577. [Abstract] [Full Text] [PDF]

				A. Hus-Citharel, N. Bouby, J. Marchetti, D. Chansel, D. Goidin, D. Gourdji, P. Corvol, and C. Llorens-Cortes Desensitization of Type 1 Angiotensin II Receptor Subtypes in the Rat Kidney Endocrinology, November 1, 2001; 142(11): 4683 - 4692. [Abstract] [Full Text] [PDF]

				A. García-Caballero, J. A. Olivares-Reyes, K. J. Catt, and J. A. García-Saínz Angiotensin AT1 Receptor Phosphorylation and Desensitization in a Hepatic Cell Line. Roles of Protein Kinase C and Phosphoinositide 3-Kinase Mol. Pharmacol., March 1, 2001; 59(3): 576 - 585. [Abstract] [Full Text]

				A. Gonzalez Iglesias, C. Suarez, C. Feierstein, G. Diaz-Torga, and D. Becu-Villalobos Desensitization of angiotensin II: effect on [Ca2+]i, inositol triphosphate, and prolactin in pituitary cells Am J Physiol Endocrinol Metab, March 1, 2001; 280(3): E462 - E470. [Abstract] [Full Text] [PDF]

				M. de Gasparo, K. J. Catt, T. Inagami, J. W. Wright, and Th. Unger International Union of Pharmacology. XXIII. The Angiotensin II Receptors Pharmacol. Rev., September 1, 2000; 52(3): 415 - 472. [Abstract] [Full Text] [PDF]

				W. G. Thomas, H. Qian, C.-S. Chang, and S. Karnik Agonist-induced Phosphorylation of the Angiotensin II (AT1A) Receptor Requires Generation of a Conformation That Is Distinct from the Inositol Phosphate-signaling State J. Biol. Chem., January 28, 2000; 275(4): 2893 - 2900. [Abstract] [Full Text] [PDF]

				Z. Xiao, Y. Yao, Y. Long, and P. Devreotes Desensitization of G-protein-coupled Receptors. AGONIST-INDUCED PHOSPHORYLATION OF THE CHEMOATTRACTANT RECEPTOR cAR1 LOWERS ITS INTRINSIC AFFINITY FOR cAMP J. Biol. Chem., January 15, 1999; 274(3): 1440 - 1448. [Abstract] [Full Text] [PDF]

				D. E. Richard, S. A. Laporte, S. G. Bernier, R. Leduc, and G. Guillemette Desensitization of AT1 Receptor-Mediated Cellular Responses Requires Long Term Receptor Down-Regulation in Bovine Adrenal Glomerulosa Cells Endocrinology, September 1, 1997; 138(9): 3828 - 3835. [Abstract] [Full Text] [PDF]

				H. Yang, D. Lu, G. P. Vinson, and M. K. Raizada Involvement of MAP Kinase in Angiotensin II-Induced Phosphorylation and Intracellular Targeting of Neuronal AT1 Receptors J. Neurosci., March 1, 1997; 17(5): 1660 - 1669. [Abstract] [Full Text] [PDF]

				M. Oppermann, N. J. Freedman, R. W. Alexander, and R. J. Lefkowitz Phosphorylation of the Type 1A Angiotensin II Receptor by G Protein-coupled Receptor Kinases and Protein Kinase C J. Biol. Chem., May 31, 1996; 271(22): 13266 - 13272. [Abstract] [Full Text] [PDF]

				M.-J. Tseng, K. Detjen, V. Struk, and C. D. Logsdon Carboxyl-terminal Domains Determine Internalization and Recycling Characteristics of Bombesin Receptor Chimeras J. Biol. Chem., August 11, 1995; 270(32): 18858 - 18864. [Abstract] [Full Text] [PDF]