3H]Benzophenone Photolabeling Identifies State-Dependent Changes in Nicotinic Acetylcholine Receptor Structure

There are no files associated with this record.

Title 3H]Benzophenone Photolabeling Identifies State-Dependent Changes in Nicotinic Acetylcholine Receptor Structure
Author III, Galo Garcia; Chiara, David C.; Nirthanan, S. Niru; Hamouda, Ayman K.; Stewart, Deidre S.; Cohen, Jonathan B.
Journal Name Biochemistry
Year Published 2007
Place of publication Washington, DC
Publisher American Chemical Society
Abstract Interactions of benzophenone (BP) with the Torpedo nicotinic acetylcholine receptor (nAChR) were characterized by electrophysiological analyses, radioligand binding assays, and photolabeling of nAChR-rich membranes with [3H]BP to identify the amino acids contributing to its binding sites. BP acted as a low potency noncompetitive antagonist, reversibly inhibiting the ACh responses of nAChRs expressed in Xenopus oocytes (IC50 = 600 μM) and the binding of the noncompetitive antagonist [3H]tetracaine to nAChR-rich membranes (IC50 = 150 μM). UV irradiation at 365 nm resulted in covalent incorporation of [3H]BP into the nAChR subunits (δ > α β > γ), with photoincorporation limited to the nAChR transmembrane domain. Comparison of nAChR photolabeling in the closed state (absence of agonist) and desensitized state (equilibrated with agonist) revealed selective desensitized state labeling in the δ subunit of δPhe-232 in δM1 and δPro-286/δIle-288 near the beginning of δM3 that are within a pocket at the interface between the transmembrane and extracellular domains. There was labeling in the closed state within the ion channel at position M2-13 (αVal-255, βVal-261, and δVal-269) that was reduced by 90% upon desensitization and labeling in the transmembrane M3 helices of the β and γ subunits (βMet-285, βMet-288, and γMet-291) that was reduced by 50−80% in the desensitized state. Labeling at the lipid interface (αMet-415 in αM4) was unaffected by agonist. These results provide a further definition of the regions in the nAChR transmembrane domain that differ in structure between the closed and desensitized states.
Peer Reviewed Yes
Published Yes
Alternative URI http://dx.doi.org/10.1021/bi7008163
Volume 46
Issue Number 36
Page from 10296
Page to 10307
ISSN 0006-2960
Date Accessioned 2009-07-09
Date Available 2009-09-28T06:50:50Z
Language en_AU
Research Centre Griffith Health Institute
Faculty Griffith Health Faculty
Subject Biochemistry and Cell Biology; Receptors and Membrane Biology
URI http://hdl.handle.net/10072/25827
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1x

Brief Record

Griffith University copyright notice