HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Habib, A. Articles by FitzGerald, G. A. Search for Related Content PubMed PubMed Citation Articles by Habib, A. Articles by FitzGerald, G. A. Social Bookmarking                      What's this?

Volume 272, Number 11, Issue of March 14, 1997 pp. 7191-7200
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Rapid, Agonist-dependent Phosphorylation in Vivo of Human Thromboxane Receptor Isoforms
MINIMAL INVOLVEMENT OF PROTEIN KINASE C

(Received for publication, October 7, 1996, and in revised form, December 5, 1996)

Aïda Habib , Roberta Vezza , Christophe Créminon ^§ , Jacques Maclouf ^¶ and Garret A. FitzGerald

From the  Center for Experimental Therapeutics, University of Pennsylvania, Philadelphia, Pennsylvania 19104, ^§ Commissariat à l'Energie Atomique (Saclay), Service de Pharmacologie et d'Immunologie, Saclay 91191 Gif-sur-Yvette, France, and ^¶ INSERM U 348, Hôpital Lariboisière, 75475 Paris Cedex 10, France

Thromboxane A₂ (TxA₂) is a potent vasoconstrictor and platelet agonist. Its biological function is tightly regulated. G protein-coupled membrane receptors transduce the effects of TxA₂. However, although a single thromboxane receptor (TP) gene has been identified, two splice variants have been cloned from human placenta and megakaryocytic lines (TP) and from human endothelial cells (TP). These differ in the length of their carboxyl-terminal extensions (15 versus 79 residues), which contain multiple potential sites for receptor phosphorylation. Given that TP agonists activate protein kinase C (PKC), it would seem possible that PKC-dependent phosphorylation of TPs might play a central role in homologous desensitization of these receptors.

To determine if the TP isoforms were differentially phosphorylated in response to agonist in vivo, human embryonic kidney (HEK) 293 cells were stably transfected with TP and TP. Isoform-specific anti-peptide antibodies were developed and used to immunoprecipitate the phosphorylated receptors. U46619, a PGH₂/TxA₂ mimetic, induced specific phosphorylation of both isoforms. Phosphorylation of the two isoforms was similar in dose and time dependence, reaching a plateau at around 100 nM U46619. Inhibition of PKC with either GF 109203X (5 µM) or RO 31-8220 (5 µM) or of protein kinase A with H-89 (50 µM) marginally influenced agonist-dependent phosphorylation of either isoform and failed to modulate homologous desensitization of agonist-induced stimulation of inositol phosphate formation. Similar results were obtained when PKC was down-regulated by long term incubation with the phorbol ester, phorbol myristate acetate. Although short term stimulation with phorbol myristate acetate caused PKC-dependent phosphorylation of TPs in vivo, thrombin stimulation of the TP-transfected HEK cells in vivo failed to phosphorylate either of the TP isoforms. Thus, despite the capacity of PKC to phosphorylate TPs in HEK 293 cells and the likely activation of PKC by TP stimulation, this enzyme, like protein kinase A, contributes marginally to rapid, agonist-induced phosphorylation of either TP isoform.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Hinton, A. Gutsol, and S. Dakshinamurti Thromboxane hypersensitivity in hypoxic pulmonary artery myocytes: altered TP receptor localization and kinetics Am J Physiol Lung Cell Mol Physiol, March 1, 2007; 292(3): L654 - L663. [Abstract] [Full Text] [PDF]

				S. J. Wilson, J. K. Dowling, L. Zhao, E. Carnish, and E. M. Smyth Regulation of Thromboxane Receptor Trafficking Through the Prostacyclin Receptor in Vascular Smooth Muscle Cells: Role of Receptor Heterodimerization Arterioscler. Thromb. Vasc. Biol., February 1, 2007; 27(2): 290 - 296. [Abstract] [Full Text] [PDF]

				C. Buccellati, A. Sala, G. Rossoni, V. Capra, G. E. Rovati, A. Di Gennaro, G. Folco, S. Colli, and C. Casagrande Pharmacological Characterization of 2NTX-99 [4-Methoxy-N1-(4-trans-nitrooxycyclohexyl)-N3-(3-pyridinylmethyl)-1,3-benzenedicarboxamide], a Potential Antiatherothrombotic Agent with Antithromboxane and Nitric Oxide Donor Activity in Platelet and Vascular Preparations J. Pharmacol. Exp. Ther., May 1, 2006; 317(2): 830 - 837. [Abstract] [Full Text] [PDF]

				E. Hamelin, C. Theriault, G. Laroche, and J.-L. Parent The Intracellular Trafficking of the G Protein-coupled Receptor TP{beta} Depends on a Direct Interaction with Rab11 J. Biol. Chem., October 28, 2005; 280(43): 36195 - 36205. [Abstract] [Full Text] [PDF]

				S. Naik, C. K. Billington, R. M. Pascual, D. A. Deshpande, F. P. Stefano, T. A. Kohout, D. M. Eckman, J. L. Benovic, and R. B. Penn Regulation of Cysteinyl Leukotriene Type 1 Receptor Internalization and Signaling J. Biol. Chem., March 11, 2005; 280(10): 8722 - 8732. [Abstract] [Full Text] [PDF]

				V. Capra, A. Veltri, C. Foglia, L. Crimaldi, A. Habib, M. Parenti, and G. E. Rovati Mutational Analysis of the Highly Conserved ERY Motif of the Thromboxane A2 Receptor: Alternative Role in G Protein-Coupled Receptor Signaling Mol. Pharmacol., October 1, 2004; 66(4): 880 - 889. [Abstract] [Full Text] [PDF]

				M. Bolla, D. You, L. Loufrani, B. I. Levy, S. Levy-Toledano, A. Habib, and D. Henrion Cyclooxygenase Involvement in Thromboxane-Dependent Contraction in Rat Mesenteric Resistance Arteries Hypertension, June 1, 2004; 43(6): 1264 - 1269. [Abstract] [Full Text] [PDF]

				H. M. Reid and B. T. Kinsella The {alpha}, but Not the {beta}, Isoform of the Human Thromboxane A2 Receptor Is a Target for Nitric Oxide-mediated Desensitization: INDEPENDENT MODULATION OF TP{alpha} SIGNALING BY NITRIC OXIDE AND PROSTACYCLIN J. Biol. Chem., December 19, 2003; 278(51): 51190 - 51202. [Abstract] [Full Text] [PDF]

				P. Nurden, C. Poujol, J. Winckler, R. Combrie, N. Pousseau, P. B. Conley, S. Levy-Toledano, A. Habib, and A. T. Nurden Immunolocalization of P2Y1 and TPalpha receptors in platelets showed a major pool associated with the membranes of alpha -granules and the open canalicular system Blood, February 15, 2003; 101(4): 1400 - 1408. [Abstract] [Full Text] [PDF]

				R. R. Price, D. P. Morris, G. Biswas, M. P. Smith, and D. A. Schwinn Acute Agonist-mediated Desensitization of the Human alpha 1a-Adrenergic Receptor Is Primarily Independent of Carboxyl Terminus Regulation. IMPLICATIONS FOR REGULATION OF alpha 1aAR SPLICE VARIANTS J. Biol. Chem., March 8, 2002; 277(11): 9570 - 9579. [Abstract] [Full Text] [PDF]

				H. Zhou, F. Yan, and H.-H. Tai Phosphorylation and Desensitization of the Human Thromboxane Receptor-alpha by G Protein-Coupled Receptor Kinases J. Pharmacol. Exp. Ther., September 1, 2001; 298(3): 1243 - 1251. [Abstract] [Full Text] [PDF]

				Z. Zhang, S. C. Austin, and E. M. Smyth Glycosylation of the Human Prostacyclin Receptor: Role in Ligand Binding and Signal Transduction Mol. Pharmacol., September 1, 2001; 60(3): 480 - 487. [Abstract] [Full Text] [PDF]

				K. P. Minneman Splice Variants of GPCRs Mol. Interv., June 1, 2001; 1(2): 108 - 116. [Abstract] [Full Text] [PDF]

				R. Vezza, J. Rokach, and G. A. FitzGerald Prostaglandin F2alpha Receptor-Dependent Regulation of Prostaglandin Transport Mol. Pharmacol., June 1, 2001; 59(6): 1506 - 1513. [Abstract] [Full Text]

				R. F. Spurney Regulation of Thromboxane Receptor (TP) Phosphorylation by Protein Phosphatase 1 (PP1) and PP2A J. Pharmacol. Exp. Ther., April 13, 2001; 296(2): 592 - 599. [Abstract] [Full Text]

				P. C. Dunlop, L. A. Leis, and G. J. Johnson Epinephrine correction of impaired platelet thromboxane receptor signaling Am J Physiol Cell Physiol, December 1, 2000; 279(6): C1760 - C1771. [Abstract] [Full Text] [PDF]

				D. Pratico, Y. Cheng, and G. A. FitzGerald TP or Not TP: Primary Mediators in a Close Runoff? Arterioscler. Thromb. Vasc. Biol., July 1, 2000; 20(7): 1695 - 1698. [Full Text] [PDF]

				L. P. Audoly, B. Rocca, J.-E. Fabre, B. H. Koller, D. Thomas, A. L. Loeb, T. M. Coffman, and G. A. FitzGerald Cardiovascular Responses to the Isoprostanes iPF2{alpha}-III and iPE2-III Are Mediated via the Thromboxane A2 Receptor In Vivo Circulation, June 20, 2000; 101(24): 2833 - 2840. [Abstract] [Full Text] [PDF]

				H. Fujino, D. Srinivasan, K. L. Pierce, and J. W. Regan Differential Regulation of Prostaglandin F2alpha Receptor Isoforms by Protein Kinase C Mol. Pharmacol., February 1, 2000; 57(2): 353 - 358. [Abstract] [Full Text]

				L. Brydon, F. Roka, L. Petit, P. de Coppet, M. Tissot, P. Barrett, P. J. Morgan, C. Nanoff, A. D. Strosberg, and R. Jockers Dual Signaling of Human Mel1a Melatonin Receptors via Gi2, Gi3, and Gq/11 Proteins Mol. Endocrinol., December 1, 1999; 13(12): 2025 - 2038. [Abstract] [Full Text]

				S. Narumiya, Y. Sugimoto, and F. Ushikubi Prostanoid Receptors: Structures, Properties, and Functions Physiol Rev, October 1, 1999; 79(4): 1193 - 1226. [Abstract] [Full Text] [PDF]

				J. M. Manganello, Y. Djellas, C. Borg, K. Antonakis, and G. C. Le Breton Cyclic AMP-dependent Phosphorylation of Thromboxane A2 Receptor-associated Galpha 13 J. Biol. Chem., September 24, 1999; 274(39): 28003 - 28010. [Abstract] [Full Text] [PDF]

				E. D. Roush, K. Warabi, and M. M. Kwatra Characterization of Differences Between Rapid Agonist-Dependent Phosphorylation and Phorbol Ester-Mediated Phosphorylation of Human Substance P Receptor in Intact Cells Mol. Pharmacol., May 1, 1999; 55(5): 855 - 862. [Abstract] [Full Text]

				R. Vezza, A. Habib, and G. A. FitzGerald Differential Signaling by the Thromboxane Receptor Isoforms via the Novel GTP-binding Protein, Gh J. Biol. Chem., April 30, 1999; 274(18): 12774 - 12779. [Abstract] [Full Text] [PDF]

				J.-L. Parent, P. Labrecque, M. J. Orsini, and J. L. Benovic Internalization of the TXA2 Receptor alpha  and beta  Isoforms. ROLE OF THE DIFFERENTIALLY SPLICED COOH TERMINUS IN AGONIST-PROMOTED RECEPTOR INTERNALIZATION J. Biol. Chem., March 26, 1999; 274(13): 8941 - 8948. [Abstract] [Full Text] [PDF]

				A. Habib, G. A. FitzGerald, and J. Maclouf Phosphorylation of the Thromboxane Receptor alpha , the Predominant Isoform Expressed in Human Platelets J. Biol. Chem., January 29, 1999; 274(5): 2645 - 2651. [Abstract] [Full Text] [PDF]

				R. F. Spurney Role of C-terminal Serines in Desensitization and Phosphorylation of the Mouse Thromboxane Receptor J. Biol. Chem., October 23, 1998; 273(43): 28496 - 28503. [Abstract] [Full Text] [PDF]

				E. M. Smyth, W. H. Li, and G. A. FitzGerald Phosphorylation of the Prostacyclin Receptor during Homologous Desensitization. A CRITICAL ROLE FOR PROTEIN KINASE C J. Biol. Chem., September 4, 1998; 273(36): 23258 - 23266. [Abstract] [Full Text] [PDF]

				M. T. Walsh, J. F. Foley, and B. T. Kinsella Characterization of the Role of N-Linked Glycosylation on the Cell Signaling and Expression of the Human Thromboxane A2 Receptor Alpha and Beta Isoforms J. Pharmacol. Exp. Ther., August 1, 1998; 286(2): 1026 - 1036. [Abstract] [Full Text]

				G. Innamorati, H. Sadeghi, and M. Birnbaumer Transient Phosphorylation of the V1a Vasopressin Receptor J. Biol. Chem., March 20, 1998; 273(12): 7155 - 7161. [Abstract] [Full Text] [PDF]

				P. Kunapuli, J. A. Lawson, J. Rokach, and G. A. FitzGerald Functional Characterization of the Ocular Prostaglandin F2alpha (PGF2alpha ) Receptor. ACTIVATION BY THE ISOPROSTANE, 12-iso-PGF2alpha J. Biol. Chem., October 24, 1997; 272(43): 27147 - 27154. [Abstract] [Full Text] [PDF]

				H. Fujino, K. L. Pierce, D. Srinivasan, C. E. Protzman, A. H. Krauss, D. F. Woodward, and J. W. Regan Delayed Reversal of Shape Change in Cells Expressing FPB Prostanoid Receptors. POSSIBLE ROLE OF RECEPTOR RESENSITIZATION J. Biol. Chem., September 15, 2000; 275(38): 29907 - 29914. [Abstract] [Full Text] [PDF]

				J.-L. Parent, P. Labrecque, M. Driss Rochdi, and J. L. Benovic Role of the Differentially Spliced Carboxyl Terminus in Thromboxane A2 Receptor Trafficking. IDENTIFICATION OF A DISTINCT MOTIF FOR TONIC INTERNALIZATION J. Biol. Chem., March 2, 2001; 276(10): 7079 - 7085. [Abstract] [Full Text] [PDF]

				M.-T. Walsh, J. F. Foley, and B. T. Kinsella The alpha , but Not the beta , Isoform of the Human Thromboxane A2 Receptor Is a Target for Prostacyclin-mediated Desensitization J. Biol. Chem., June 30, 2000; 275(27): 20412 - 20423. [Abstract] [Full Text] [PDF]