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

J. Biol. Chem., Vol. 280, Issue 34, 30460-30468, August 26, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/34/30460    most recent M504229200v1 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 Dutertre, S. Articles by Lewis, R. J. Search for Related Content PubMed PubMed Citation Articles by Dutertre, S. Articles by Lewis, R. J. Social Bookmarking                      What's this?

2 Subunit Contribution to 4/7 -Conotoxin Binding to the Nicotinic Acetylcholine Receptor^*

Sébastien Dutertre, Annette Nicke¶||, and Richard J. Lewis**

From the Institute for Molecular Bioscience, The University of Queensland, Queensland 4072, Australia and the ^¶Max Planck Institute for Experimental Medicine, Hermann Rein-Strasse 3, 37075 Göttingen, Germany

The structures of acetylcholine-binding protein (AChBP) and nicotinic acetylcholine receptor (nAChR) homology models have been used to interpret data from mutagenesis experiments at the nAChR. However, little is known about AChBP-derived structures as predictive tools. Molecular surface analysis of nAChR models has revealed a conserved cleft as the likely binding site for the 4/7 -conotoxins. Here, we used an 32 model to identify 2 subunit residues in this cleft and investigated their influence on the binding of -conotoxins MII, PnIA, and GID to the 32 nAChR by two-electrode voltage clamp analysis. Although a 2-L119Q mutation strongly reduced the affinity of all three -conotoxins, 2-F117A, 2-V109A, and 2-V109G mutations selectively enhanced the binding of MII and GID. An increased activity of -conotoxins GID and MII was also observed when the 2-F117A mutant was combined with the 4 instead of the 3 subunit. Investigation of A10L-PnIA indicated that high affinity binding to 2-F117A, 2-V109A, and 2-V109G mutants was conferred by amino acids with a long side chain in position 10 (PnIA numbering). Docking simulations of 4/7 -conotoxin binding to the 32 model supported a direct interaction between mutated nAChR residues and -conotoxin residues 6, 7, and 10. Taken together, these data provide evidence that the subunit contributes to -conotoxin binding and selectivity and demonstrate that a small cleft leading to the agonist binding site is targeted by -conotoxins to block the nAChR.

Received for publication, April 19, 2005 , and in revised form, June 1, 2005.

^* This work was supported by Grant DP0208295 from the Australian Research Council, Grant NI 592/3-1 from the Deutsche Forschungsgemeinschaft, and a postgraduate scholarship from The University of Queensland (to S. D.). 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.

These authors contributed equally to this work.

^|| Current address: Max Planck Institute for Brain Research, Deutschordenstr. 46, D-60528 Frankfurt, Germany.

^** To whom correspondence should be addressed: Inst. for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia. Tel.: 61-7-3346-2984; Fax: 61-7-3346-2101; E-mail: r.lewis{at}imb.uq.edu.au.

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

This article has been cited by other articles:

				I. Bartholomaus, L. Milan-Lobo, A. Nicke, S. Dutertre, H. Hastrup, A. Jha, U. Gether, H. H. Sitte, H. Betz, and V. Eulenburg Glycine Transporter Dimers: EVIDENCE FOR OCCURRENCE IN THE PLASMA MEMBRANE J. Biol. Chem., April 18, 2008; 283(16): 10978 - 10991. [Abstract] [Full Text] [PDF]

				E. JERLHAG, M. GROTLI, K. LUTHMAN, L. SVENSSON, and J. A. ENGEL ROLE OF THE SUBUNIT COMPOSITION OF CENTRAL NICOTINIC ACETYLCHOLINE RECEPTORS FOR THE STIMULATORY AND DOPAMINE-ENHANCING EFFECTS OF ETHANOL Alcohol Alcohol., September 1, 2006; 41(5): 486 - 493. [Abstract] [Full Text] [PDF]

				M. Loughnan, A. Nicke, A. Jones, C. I. Schroeder, S. T. Nevin, D. J. Adams, P. F. Alewood, and R. J. Lewis Identification of a Novel Class of Nicotinic Receptor Antagonists: DIMERIC CONOTOXINS VxXIIA, VxXIIB, and VxXIIC FROM CONUS VEXILLUM J. Biol. Chem., August 25, 2006; 281(34): 24745 - 24755. [Abstract] [Full Text] [PDF]

				T. T. Talley, B. M. Olivera, K.-H. Han, S. B. Christensen, C. Dowell, I. Tsigelny, K.-Y. Ho, P. Taylor, and J. M. McIntosh {alpha}-Conotoxin OmIA Is a Potent Ligand for the Acetylcholine-binding Protein as Well as {alpha}3beta2 and {alpha}7 Nicotinic Acetylcholine Receptors J. Biol. Chem., August 25, 2006; 281(34): 24678 - 24686. [Abstract] [Full Text] [PDF]

				R. J. Clark, H. Fischer, S. T. Nevin, D. J. Adams, and D. J. Craik The Synthesis, Structural Characterization, and Receptor Specificity of the {alpha}-Conotoxin Vc1.1 J. Biol. Chem., August 11, 2006; 281(32): 23254 - 23263. [Abstract] [Full Text] [PDF]

				C. Ulens, R. C. Hogg, P. H. Celie, D. Bertrand, V. Tsetlin, A. B. Smit, and T. K. Sixma Structural determinants of selective {alpha}-conotoxin binding to a nicotinic acetylcholine receptor homolog AChBP PNAS, March 7, 2006; 103(10): 3615 - 3620. [Abstract] [Full Text] [PDF]