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

J. Biol. Chem., Vol. 275, Issue 48, 37860-37869, December 1, 2000

This Article Full Text Full Text (PDF) All Versions of this Article: 275/48/37860    most recent M006709200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Schulz, A. Articles by Schöneberg, T. Search for Related Content PubMed PubMed Citation Articles by Schulz, A. Articles by Schöneberg, T. Social Bookmarking                      What's this?

Requirement of Specific Intrahelical Interactions for Stabilizing the Inactive Conformation of Glycoprotein Hormone Receptors^*

Angela Schulz, Karsten Bruns^§, Peter Henklein^¶, Gerd Krause, Mario Schubert, Thomas Gudermann^**, Victor Wray^§, Günter Schultz, and Torsten Schöneberg

From the  Institut für Pharmakologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Thielallee 69-73, 14195 Berlin, Germany, the ^§ Department of Molecular Structure Research, Gesellschaft für Biotechnologische Forschung, Braunschweig, Mascheroder Weg 1, 38124 Braunschweig, Germany, the ^¶ Institut für Biochemie, Medizinische Fakultät (Charité) der Humboldt-Universität zu Berlin, Schumannstrasse 20/21, 10098 Berlin, Germany, the  Institut für Molekulare Pharmakologie, Alfred Kowalke Strasse 4, 10315 Berlin, Germany, and the ^** Institut für Pharmakologie und Toxikologie, Fachbereich Humanmedizin, Philipps-Universität Marburg, Karl-von-Frisch-Strasse 1, 35033 Marburg, Germany

Systematic analysis of structural changes induced by activating mutations has been frequently utilized to study activation mechanisms of G-protein-coupled receptors (GPCRs). In the thyrotropin receptor and the lutropin receptor (LHR), a large number of naturally occurring mutations leading to constitutive receptor activation were identified. Saturating mutagenesis studies of a highly conserved Asp in the junction of the third intracellular loop and transmembrane domain 6 suggested a participation of this anionic residue in a salt bridge stabilizing the inactive receptor conformation. However, substitution of all conserved cationic residues at the cytoplasmic receptor surface did not support this hypothesis. Asp/Glu residues are a common motif at the N-terminal ends of -helices terminating and stabilizing the helical structure (helix capping). Since Asp/Glu residues in the third intracellular loop/transmembrane domain 6 junction are not only preserved in glycoprotein hormone receptors but also in other GPCRs we speculated that this residue probably participates in an N-terminal helix-capping structure. Poly-Ala stretches are known to form and stabilize -helices. Herein, we show that the function of the highly conserved Asp can be mimicked by poly-Ala substitutions in the LHR and thyrotropin receptor. CD and NMR studies of peptides derived from the juxtamembrane portion of the LHR confirmed the helix extension by the poly-Ala substitution and provided further evidence for an involvement of Asp in a helix-capping structure. Our data implicate that in addition to well established interhelical interactions the inactive conformation of GPCRs is also stabilized by specific intrahelical structures.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				M. Zhong, B. Parish, D. A. Murtazina, C.-Y. Ku, and B. M. Sanborn Amino acids in the COOH-terminal region of the oxytocin receptor third intracellular domain are important for receptor function Am J Physiol Endocrinol Metab, April 1, 2007; 292(4): E977 - E984. [Abstract] [Full Text] [PDF]

				B. Karges, G. Krause, J. Homoki, K.-M. Debatin, N. de Roux, and W. Karges TSH receptor mutation V509A causes familial hyperthyroidism by release of interhelical constraints between transmembrane helices TMH3 and TMH5 J. Endocrinol., August 1, 2005; 186(2): 377 - 385. [Abstract] [Full Text] [PDF]

				K. Sangkuhl, A. Schulz, G. Schultz, and T. Schoneberg Structural Requirements for Mutational Lutropin/Choriogonadotropin Receptor Activation J. Biol. Chem., November 27, 2002; 277(49): 47748 - 47755. [Abstract] [Full Text] [PDF]

				K. Angelova, F. Fanelli, and D. Puett A Model for Constitutive Lutropin Receptor Activation Based on Molecular Simulation and Engineered Mutations in Transmembrane Helices 6 and 7 J. Biol. Chem., August 23, 2002; 277(35): 32202 - 32213. [Abstract] [Full Text] [PDF]

				S. Mukherjee, V. V. Gurevich, A. Preninger, H. E. Hamm, M.-F. Bader, A. T. Fazleabas, L. Birnbaumer, and M. Hunzicker-Dunn Aspartic Acid 564 in the Third Cytoplasmic Loop of the Luteinizing Hormone/Choriogonadotropin Receptor Is Crucial for Phosphorylation-independent Interaction with Arrestin2 J. Biol. Chem., May 10, 2002; 277(20): 17916 - 17927. [Abstract] [Full Text] [PDF]

				M. Waelbroeck Activation of Guanosine 5'-[{gamma}-35S]thio-triphosphate Binding through M1 Muscarinic Receptors in Transfected Chinese Hamster Ovary Cell Membranes: 2. Testing the "Two-States" Model of Receptor Activation Mol. Pharmacol., April 1, 2001; 59(4): 886 - 893. [Abstract] [Full Text]