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 Marjamäki, A. Articles by Scheinin, M. Search for Related Content PubMed PubMed Citation Articles by Marjamäki, A. Articles by Scheinin, M. Social Bookmarking                      What's this?

J Biol Chem, Vol. 274, Issue 31, 21867-21872, July 30, 1999

Chloroethylclonidine and 2-Aminoethyl Methanethiosulfonate Recognize Two Different Conformations of the Human _2A-Adrenergic Receptor

Anne Marjamäki, Heini Frang, Marjo Pihlavisto, Anna-Marja Hoffrén^¶, Tiina Salminen, Mark S. Johnson, Jaana Kallio, Jonathan A. Javitch^**, and Mika Scheinin

From the  Department of Pharmacology and Clinical Pharmacology, University of Turku, MediCity, ^¶ Juvantia Pharma Ltd, and  Department of Biochemistry and Pharmacy, Åbo Akademi University and Turku Center for Biotechnology, University of Turku and Åbo Akademi University, Tykistökatu 6 A, FIN-20520 Turku, Finland and ^** Center for Molecular Recognition and the Departments of Psychiatry and Pharmacology, Columbia University College of Physicians and Surgeons, New York, New York 10032

The substituted cysteine-accessibility method and two sulfhydryl-specific reagents, the methane-thiosulfonate derivative 2-aminoethyl methanethiosulfonate (MTSEA) and the ₂-adrenergic receptor (₂-AR) agonist chloroethylclonidine (CEC), were used to determine the relative accessibility of engineered cysteines in the fifth transmembrane domain of the human _2A-AR (H2A). The second-order rate constants for the reaction of the receptor with MTSEA and CEC were determined with the wild type H2A (cysteine at position 201) and receptor mutants containing accessible cysteines at other positions within the binding-site crevice (positions 197, 200, and 204). The rate of reaction of CEC was similar to that of MTSEA at residues Cys-197, Cys-201, and Cys-204. The rate of reaction of CEC with Cys-200, however, was more than 5 times that of MTSEA, suggesting that these compounds may interact with two different receptor conformations. MTSEA, having no recognition specificity for the receptor, likely reacts with the predominant inactive receptor conformation (R), whereas the agonist CEC may stabilize and react preferentially with the active receptor conformation (R*). This hypothesis was consistent with three-dimensional receptor-ligand models, which further suggest that _2A-AR activation may involve the clockwise rotation of transmembrane domain 5.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Zurn, U. Zabel, J.-P. Vilardaga, H. Schindelin, M. J. Lohse, and C. Hoffmann Fluorescence Resonance Energy Transfer Analysis of {alpha}2a-Adrenergic Receptor Activation Reveals Distinct Agonist-Specific Conformational Changes Mol. Pharmacol., March 1, 2009; 75(3): 534 - 541. [Abstract] [Full Text] [PDF]

				G. Liapakis, W. C. Chan, M. Papadokostaki, and J. A. Javitch Synergistic Contributions of the Functional Groups of Epinephrine to Its Affinity and Efficacy at the {beta}2 Adrenergic Receptor Mol. Pharmacol., May 1, 2004; 65(5): 1181 - 1190. [Abstract] [Full Text]

				P. J. Pauwels, I. Rauly, and T. Wurch Dissimilar Pharmacological Responses by a New Series of Imidazoline Derivatives at Precoupled and Ligand-Activated {alpha}2A-Adrenoceptor States: Evidence for Effector Pathway-Dependent Differential Antagonism J. Pharmacol. Exp. Ther., June 1, 2003; 305(3): 1015 - 1023. [Abstract] [Full Text] [PDF]

				C. Govaerts, A. Bondue, J.-Y. Springael, M. Olivella, X. Deupi, E. Le Poul, S. J. Wodak, M. Parmentier, L. Pardo, and C. Blanpain Activation of CCR5 by Chemokines Involves an Aromatic Cluster between Transmembrane Helices 2 and 3 J. Biol. Chem., January 10, 2003; 278(3): 1892 - 1903. [Abstract] [Full Text] [PDF]

				T. Nyrönen, M. Pihlavisto, J. M. Peltonen, A.-M. Hoffrén, M. Varis, T. Salminen, S. Wurster, A. Marjamäki, L. Kanerva, E. Katainen, et al. Molecular Mechanism for Agonist-Promoted alpha 2A-Adrenoceptor Activation by Norepinephrine and Epinephrine Mol. Pharmacol., April 16, 2001; 59(5): 1343 - 1354. [Abstract] [Full Text]

				D. A. Shapiro, K. Kristiansen, W. K. Kroeze, and B. L. Roth Differential Modes of Agonist Binding to 5-Hydroxytryptamine2A Serotonin Receptors Revealed by Mutation and Molecular Modeling of Conserved Residues in Transmembrane Region 5 Mol. Pharmacol., November 1, 2000; 58(5): 877 - 886. [Abstract] [Full Text]

				C. M. Topham, L. Mouledous, and J.-C. Meunier On the spatial disposition of the fifth transmembrane helix and the structural integrity of the transmembrane binding site in the opioid and ORL1 G protein-coupled receptor family Protein Eng. Des. Sel., July 1, 2000; 13(7): 477 - 490. [Abstract] [Full Text] [PDF]

				K. Allman, K. M. Page, C. A.M. Curtis, and E. C. Hulme Scanning Mutagenesis Identifies Amino Acid Side Chains in Transmembrane Domain 5 of the M1 Muscarinic Receptor that Participate in Binding the Acetyl Methyl Group of Acetylcholine Mol. Pharmacol., July 1, 2000; 58(1): 175 - 184. [Abstract] [Full Text]

				T. Salminen, M. Varis, T. Nyronen, M. Pihlavisto, A.-M. Hoffren, T. Lonnberg, A. Marjamaki, H. Frang, J.-M. Savola, M. Scheinin, et al. Three-dimensional Models of alpha 2A-Adrenergic Receptor Complexes Provide a Structural Explanation for Ligand Binding J. Biol. Chem., August 13, 1999; 274(33): 23405 - 23413. [Abstract] [Full Text] [PDF]

				H. Frang, V. Cockcroft, T. Karskela, M. Scheinin, and A. Marjamaki Phenoxybenzamine Binding Reveals the Helical Orientation of the Third Transmembrane Domain of Adrenergic Receptors J. Biol. Chem., August 10, 2001; 276(33): 31279 - 31284. [Abstract] [Full Text] [PDF]