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 Prossnitz, E. R. Search for Related Content PubMed PubMed Citation Articles by Prossnitz, E. R. Social Bookmarking                      What's this?

Volume 272, Number 24, Issue of June 13, 1997 pp. 15213-15219
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Desensitization of N-Formylpeptide Receptor-mediated Activation Is Dependent upon Receptor Phosphorylation

(Received for publication, October 8, 1996, and in revised form, March 21, 1997)

From the Department of Immunology, Scripps Research Institute, La Jolla, California 92037

The human N-formylpeptide receptor (FPR) represents one of the most thoroughly studied leukocyte chemoattractant receptors. Despite this, little is known about the molecular mechanisms involved in the activation and desensitization of this receptor. To assess the role of phosphorylation in receptor function, U937 promonocytic cells were stably transfected to express the recombinant human FPR. Three mutant forms of the FPR lacking specific serine and threonine residues in the receptor C terminus were studied with respect to activation and desensitization. Replacement of all 11 serine and threonine residues within the C terminus by alanine and glycine residues (ST) resulted in a receptor capable of ligand binding and G protein activation similar to the wild-type receptor. However, whereas the wild-type FPR was phosphorylated on both serine and threonine residues upon exposure to agonist and displayed a significantly reduced ability to stimulate G protein-mediated GTP hydrolysis upon subsequent exposure to agonist, ST demonstrated a complete lack of phosphorylation and displayed little alteration in its ability to stimulate G protein-mediated GTP hydrolysis upon a subsequent exposure to agonist. In addition to desensitization of G protein-mediated GTP hydrolysis, calcium mobilization was assayed to test whether desensitization occurred at a site distal to G protein activation. However, as observed with G protein activation, ST underwent no desensitization of the calcium mobilization response upon a second exposure to agonist. To define more precisely the role of specific serine and threonine residues, two additional mutants were analyzed. Replacement either of Ser³²⁸, Thr³²⁹, Thr³³¹, and Ser³³² (mutant A) or of Thr³³⁴, Thr³³⁶, Ser³³⁸, and Thr³³⁹ (mutant B) resulted in functional receptors that exhibited ~50% the level of phosphorylation following stimulation. Whereas mutant A, like ST, could not be significantly desensitized by exposure to agonist, mutant B exhibited partial desensitization. These results indicate that phosphorylation of the FPR is a necessary and sufficient step in cellular desensitization, that multiple phosphorylation sites are involved, and that redundant desensitization does not occur downstream of G protein activation in the signaling cascade.

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

This article has been cited by other articles:

				M. Riesselman, H. M. Miettinen, J. M. Gripentrog, C. I. Lord, B. Mumey, E. A. Dratz, J. Stie, R. M. Taylor, and A. J. Jesaitis C-Terminal Tail Phosphorylation of N-Formyl Peptide Receptor: Differential Recognition of Two Neutrophil Chemoattractant Receptors by Monoclonal Antibodies NFPR1 and NFPR2 J. Immunol., August 15, 2007; 179(4): 2520 - 2531. [Abstract] [Full Text] [PDF]

				A. Chigaev, A. Waller, G. J. Zwartz, T. Buranda, and L. A. Sklar Regulation of Cell Adhesion by Affinity and Conformational Unbending of {alpha}4beta1 Integrin J. Immunol., June 1, 2007; 178(11): 6828 - 6839. [Abstract] [Full Text] [PDF]

				M. Xue, G. Hsieh, M. A. Raymond-Stintz, J. Pfeiffer, D. Roberts, S. L. Steinberg, J. M. Oliver, E. R. Prossnitz, D. S. Lidke, and B. S. Wilson Activated N-Formyl Peptide Receptor and High-Affinity IgE Receptor Occupy Common Domains for Signaling and Internalization Mol. Biol. Cell, April 1, 2007; 18(4): 1410 - 1420. [Abstract] [Full Text] [PDF]

				R. M. Potter, D. C. Maestas, D. F. Cimino, and E. R. Prossnitz Regulation of N-Formyl Peptide Receptor Signaling and Trafficking by Individual Carboxyl-Terminal Serine and Threonine Residues J. Immunol., May 1, 2006; 176(9): 5418 - 5425. [Abstract] [Full Text] [PDF]

				B. S. Edwards, C. Bologa, S. M. Young, K. V. Balakin, E. R. Prossnitz, N. P. Savchuck, L. A. Sklar, and T. I. Oprea Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists Mol. Pharmacol., November 1, 2005; 68(5): 1301 - 1310. [Abstract] [Full Text] [PDF]

				S. M. Young, C. Bologa, E. R. Prossnitz, T. I. Oprea, L. A. Sklar, and B. S. Edwards High-Throughput Screening with HyperCyt(R) Flow Cytometry to Detect Small Molecule Formylpeptide Receptor Ligands J Biomol Screen, June 1, 2005; 10(4): 374 - 382. [Abstract] [PDF]

				M. Xue, C. M. Vines, T. Buranda, D. F. Cimino, T. A. Bennett, and E. R. Prossnitz N-Formyl Peptide Receptors Cluster in an Active Raft-associated State Prior to Phosphorylation J. Biol. Chem., October 22, 2004; 279(43): 45175 - 45184. [Abstract] [Full Text] [PDF]

				C. Weiss-Haljiti, C. Pasquali, H. Ji, C. Gillieron, C. Chabert, M.-L. Curchod, E. Hirsch, A. J. Ridley, R. H. van Huijsduijnen, M. Camps, et al. Involvement of Phosphoinositide 3-Kinase {gamma}, Rac, and PAK Signaling in Chemokine-induced Macrophage Migration J. Biol. Chem., October 8, 2004; 279(41): 43273 - 43284. [Abstract] [Full Text] [PDF]

				A. Chigaev, G. J. Zwartz, T. Buranda, B. S. Edwards, E. R. Prossnitz, and L. A. Sklar Conformational Regulation of {alpha}4{beta}1-Integrin Affinity by Reducing Agents: "INSIDE-OUT" SIGNALING IS INDEPENDENT OF AND ADDITIVE TO REDUCTION-REGULATED INTEGRIN ACTIVATION J. Biol. Chem., July 30, 2004; 279(31): 32435 - 32443. [Abstract] [Full Text] [PDF]

				C. M. Revankar, C. M. Vines, D. F. Cimino, and E. R. Prossnitz Arrestins Block G Protein-coupled Receptor-mediated Apoptosis J. Biol. Chem., June 4, 2004; 279(23): 24578 - 24584. [Abstract] [Full Text] [PDF]

				S. M. Young, M. S. Curry, J. T. Ransom, J. A. Ballesteros, E. R. Prossnitz, L. A. Sklar, and B. S. Edwards High-Throughput Microfluidic Mixing and Multiparametric Cell Sorting for Bioactive Compound Screening J Biomol Screen, March 1, 2004; 9(2): 103 - 111. [Abstract] [PDF]

				C. M. Vines, M. Xue, D. C. Maestas, D. F. Cimino, and E. R. Prossnitz Regulation of N-Formyl Peptide-Mediated Degranulation by Receptor Phosphorylation J. Immunol., December 15, 2002; 169(12): 6760 - 6766. [Abstract] [Full Text] [PDF]

				T. E. Graham, E. R. Prossnitz, and R. I. Dorin Dexras1/AGS-1 Inhibits Signal Transduction from the Gi-coupled Formyl Peptide Receptor to Erk-1/2 MAP Kinases J. Biol. Chem., March 22, 2002; 277(13): 10876 - 10882. [Abstract] [Full Text] [PDF]

				R. M. Potter, T. A. Key, V. V. Gurevich, L. A. Sklar, and E. R. Prossnitz Arrestin Variants Display Differential Binding Characteristics for the Phosphorylated N-Formyl Peptide Receptor Carboxyl Terminus J. Biol. Chem., March 8, 2002; 277(11): 8970 - 8978. [Abstract] [Full Text] [PDF]

				D. C. Maestes, R. M. Potter, and E. R. Prossnitz Differential Phosphorylation Paradigms Dictate Desensitization and Internalization of the N-Formyl Peptide Receptor J. Biol. Chem., October 15, 1999; 274(42): 29791 - 29795. [Abstract] [Full Text] [PDF]

				G. R. Sambrano and S. R. Coughlin The Carboxyl Tail of Protease-activated Receptor-1 Is Required for Chemotaxis. CORRELATION OF SIGNAL TERMINATION AND DIRECTIONAL MIGRATION J. Biol. Chem., July 16, 1999; 274(29): 20178 - 20184. [Abstract] [Full Text] [PDF]

				M. Bunemann and M M. Hosey G-protein coupled receptor kinases as modulators of G-protein signalling J. Physiol., May 15, 1999; 517(1): 5 - 23. [Abstract] [Full Text] [PDF]

				M. J. Rane, J. M. Arthur, E. R. Prossnitz, and K. R. McLeish Activation of Mitogen-activated Protein Kinases by Formyl Peptide Receptors Is Regulated by the Cytoplasmic Tail J. Biol. Chem., August 14, 1998; 273(33): 20916 - 20923. [Abstract] [Full Text] [PDF]

				R. M. Richardson, H. Ali, B. C. Pridgen, B. Haribabu, and R. Snyderman Multiple Signaling Pathways of Human Interleukin-8 Receptor A. INDEPENDENT REGULATION BY PHOSPHORYLATION J. Biol. Chem., April 24, 1998; 273(17): 10690 - 10695. [Abstract] [Full Text] [PDF]

				M. H. Hsu, S. C. Chiang, R. D. Ye, and E. R. Prossnitz Phosphorylation of the N-Formyl Peptide Receptor Is Required for Receptor Internalization but Not Chemotaxis J. Biol. Chem., November 21, 1997; 272(47): 29426 - 29429. [Abstract] [Full Text] [PDF]

				T. A. Bennett, D. C. Maestas, and E. R. Prossnitz Arrestin Binding to the G Protein-coupled N-Formyl Peptide Receptor Is Regulated by the Conserved "DRY" Sequence J. Biol. Chem., August 4, 2000; 275(32): 24590 - 24594. [Abstract] [Full Text] [PDF]

				J. R. Backstrom, R. D. Price, D. T. Reasoner, and E. Sanders-Bush Deletion of the Serotonin 5-HT2C Receptor PDZ Recognition Motif Prevents Receptor Phosphorylation and Delays Resensitization of Receptor Responses J. Biol. Chem., July 28, 2000; 275(31): 23620 - 23626. [Abstract] [Full Text] [PDF]

				A. Chigaev, A. M. Blenc, J. V. Braaten, N. Kumaraswamy, C. L. Kepley, R. P. Andrews, J. M. Oliver, B. S. Edwards, E. R. Prossnitz, R. S. Larson, et al. Real Time Analysis of the Affinity Regulation of alpha 4-Integrin. THE PHYSIOLOGICALLY ACTIVATED RECEPTOR IS INTERMEDIATE IN AFFINITY BETWEEN RESTING AND Mn2+ OR ANTIBODY ACTIVATION J. Biol. Chem., December 21, 2001; 276(52): 48670 - 48678. [Abstract] [Full Text] [PDF]