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

J. Biol. Chem., Vol. 279, Issue 30, 31177-31182, July 23, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/30/31177    most recent M404113200v1 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?

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^*

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, which contains numerous potentially important drug targets. Delineation of themes for agonist binding and activation of these receptors will facilitate the rational design of receptor-active drugs. We reported previously that a photolabile residue within the carboxyl-terminal half (residue 26) and mid-region (residue 16) of calcitonin covalently label the extracellular amino-terminal domain of this receptor (Dong, M., Pinon, D. I., Cox, R. F., and Miller, L. J. (2004) J. Biol. Chem. 279, 1167–1175). Chimeric receptor studies support the importance of this region and suggest important contributions of extracellular loop domains. To examine whether other parts of the ligand may contact those loops, we developed another probe that has its photolabile site of labeling within the amino-terminal half in position 8 of the ligand. This probe was a full agonist (EC₅₀ = 563 ± 67 pM), stimulating cAMP accumulation in receptor-bearing human embryonic kidney 293 cells in a concentration-dependent manner. It bound specifically and saturably (K_i = 14.3 ± 1.9 nM) and was able to efficiently label the calcitonin receptor. By purification, specific cleavage, and sequencing of labeled wild-type and mutant calcitonin receptors, the site of attachment was identified as residue Leu³⁶⁸ within the third extracellular loop of the receptor, a domain distinct from that labeled by previous probes. These data are consistent with a common ligand binding mechanism for receptors in this important family.

Received for publication, April 13, 2004 , and in revised form, May 19, 2004.

^* 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:

				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]

				M. Dong, F. Gao, D. I. Pinon, and L. J. Miller Insights into the Structural Basis of Endogenous Agonist Activation of Family B G Protein-Coupled Receptors Mol. Endocrinol., June 1, 2008; 22(6): 1489 - 1499. [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, K. Hosohata, D. I. Pinon, N. Muthukumaraswamy, and L. J. Miller Differential Spatial Approximation between Secretin and Its Receptor Residues in Active and Inactive Conformations Demonstrated by Photoaffinity Labeling Mol. Endocrinol., July 1, 2006; 20(7): 1688 - 1698. [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]

				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]