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

J. Biol. Chem., Vol. 279, Issue 2, 1167-1175, January 9, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/2/1167    most recent M305719200v1 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 Dong, M. Articles by Miller, L. J. Search for Related Content PubMed PubMed Citation Articles by Dong, M. Articles by Miller, L. J. Social Bookmarking                      What's this?

Importance of the Amino Terminus in Secretin Family G Protein-coupled Receptors

INTRINSIC PHOTOAFFINITY LABELING ESTABLISHES INITIAL DOCKING CONSTRAINTS FOR THE CALCITONIN RECEPTOR^*

Maoqing Dong, Delia I. Pinon, Richard F. Cox, and Laurence J. Miller

From the Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic Scottsdale, Scottsdale, Arizona 85259 and GlaxoSmithKline, Research Triangle Park, North Carolina 27709

The calcitonin receptor is a member of the class B family of G protein-coupled receptors, closely related to secretin and parathyroid hormone receptors. Although mechanisms of ligand binding have been directly explored for those receptors, current knowledge of the molecular basis of calcitonin binding to its receptor is based only on receptor mutagenesis. In this work we have utilized the more direct approach of photoaffinity labeling to explore spatial approximations between distinct residues within calcitonin and its receptor. For this we have developed two human calcitonin analogues incorporating a photolabile p-benzoyl-L-phenylalanine residue in the mid-region and carboxyl-terminal half of the peptide in positions 16 and 26, respectively. Both probes specifically bound to the human calcitonin receptor with high affinity and were potent stimulants of cAMP accumulation in calcitonin receptor-bearing human embryonic kidney 293 cells. They covalently labeled the calcitonin receptor in a saturable and specific manner. Further purification, deglycosylation, specific chemical and enzymatic cleavage, and sequencing of labeled wild type and mutant calcitonin receptors identified the sites of labeling for the position 16 and 26 probes as receptor residues Phe¹³⁷ and Thr³⁰, respectively. Both were within the extracellular amino terminus of the calcitonin receptor, with the former adjacent to the first transmembrane segment and the latter within the distal amino-terminal tail of the receptor. These data are consistent with affinity labeling of other members of the class B G protein-coupled receptors using analogous probes and may suggest a common ligand binding mechanism for this family.

Received for publication, June 2, 2003 , and in revised form, October 27, 2003.

^* This work was supported by a grant from GlaxoSmithKline and by National Institutes of Health Grant DK46577 (to L. J. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Mayo Clinic Scottsdale, Johnson Research Bldg., 13400 E. Shea Blvd., Scottsdale, AZ 85259. Tel.: 480-301-6830; Fax: 480-301-9162; E-mail: dongmq{at}mayo.edu.

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

This article has been cited by other articles:

				I. Assil-Kishawi, T. A. Samra, D. F. Mierke, and A. B. Abou-Samra Residue 17 of Sauvagine Cross-links to the First Transmembrane Domain of Corticotropin-releasing Factor Receptor 1 (CRFR1) J. Biol. Chem., December 19, 2008; 283(51): 35644 - 35651. [Abstract] [Full Text] [PDF]

				M. Dong, P. C.-H. Lam, D. I. Pinon, P. M. Sexton, R. Abagyan, and L. J. Miller Spatial Approximation between Secretin Residue Five and the Third Extracellular Loop of Its Receptor Provides New Insight into the Molecular Basis of Natural Agonist Binding Mol. Pharmacol., August 1, 2008; 74(2): 413 - 422. [Abstract] [Full Text] [PDF]

				K. G. Harikumar, P. C.-H. Lam, M. Dong, P. M. Sexton, R. Abagyan, and L. J. Miller Fluorescence Resonance Energy Transfer Analysis of Secretin Docking to Its Receptor: MAPPING DISTANCES BETWEEN RESIDUES DISTRIBUTED THROUGHOUT THE LIGAND PHARMACOPHORE AND DISTINCT RECEPTOR RESIDUES J. Biol. Chem., November 9, 2007; 282(45): 32834 - 32843. [Abstract] [Full Text] [PDF]

				G. Andreotti, B. L. Mendez, P. Amodeo, M. A. C. Morelli, H. Nakamuta, and A. Motta Structural Determinants of Salmon Calcitonin Bioactivity: THE ROLE OF THE LEU-BASED AMPHIPATHIC {alpha}-HELIX J. Biol. Chem., August 25, 2006; 281(34): 24193 - 24203. [Abstract] [Full Text] [PDF]

				M. Dong, D. I. Pinon, Y. W. Asmann, and L. J. Miller Possible Endogenous Agonist Mechanism for the Activation of Secretin Family G Protein-Coupled Receptors Mol. Pharmacol., July 1, 2006; 70(1): 206 - 213. [Abstract] [Full Text] [PDF]

				T. Dean, A. Linglart, M. J. Mahon, M. Bastepe, H. Juppner, J. T. Potts Jr., and T. J. Gardella Mechanisms of Ligand Binding to the Parathyroid Hormone (PTH)/PTH-Related Protein Receptor: Selectivity of a Modified PTH(1-15) Radioligand for G{alpha}S-Coupled Receptor Conformations Mol. Endocrinol., April 1, 2006; 20(4): 931 - 943. [Abstract] [Full Text] [PDF]

				V. Pham, M. Dong, J. D. Wade, L. J. Miller, C. J. Morton, H.-l. Ng, M. W. Parker, and P. M. Sexton Insights into Interactions between the {alpha}-Helical Region of the Salmon Calcitonin Antagonists and the Human Calcitonin Receptor using Photoaffinity Labeling J. Biol. Chem., August 5, 2005; 280(31): 28610 - 28622. [Abstract] [Full Text] [PDF]

				M. Dong, D. I. Pinon, and L. J. Miller Insights into the Structure and Molecular Basis of Ligand Docking to the G Protein-Coupled Secretin Receptor Using Charge-Modified Amino-Terminal Agonist Probes Mol. Endocrinol., July 1, 2005; 19(7): 1821 - 1836. [Abstract] [Full Text] [PDF]

				N. Shimizu, T. Dean, J. C. Tsang, A. Khatri, J. T Potts Jr, and T. J. Gardella Novel Parathyroid Hormone (PTH) Antagonists That Bind to the Juxtamembrane Portion of the PTH/PTH-related Protein Receptor J. Biol. Chem., January 21, 2005; 280(3): 1797 - 1807. [Abstract] [Full Text] [PDF]

				M. Dong, D. I. Pinon, R. F. Cox, and L. J. Miller Molecular Approximation between a Residue in the Amino-terminal Region of Calcitonin and the Third Extracellular Loop of the Class B G Protein-coupled Calcitonin Receptor J. Biol. Chem., July 23, 2004; 279(30): 31177 - 31182. [Abstract] [Full Text] [PDF]

				D. Koller, L. M. Ittner, R. Muff, K. Husmann, J. A. Fischer, and W. Born Selective Inactivation of Adrenomedullin over Calcitonin Gene-related Peptide Receptor Function by the Deletion of Amino Acids 14-20 of the Mouse Calcitonin-like Receptor J. Biol. Chem., May 7, 2004; 279(19): 20387 - 20391. [Abstract] [Full Text] [PDF]