期刊
BIOCHEMISTRY
卷 46, 期 36, 页码 10296-10307出版社
AMER CHEMICAL SOC
DOI: 10.1021/bi7008163
关键词
-
资金
- NIGMS NIH HHS [GM-58448] Funding Source: Medline
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 [H-3]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,mu M) and the binding of the noncompetitive antagonist [3H]-tetracaine to nAChR-rich membranes (IC50 = 150 mu M). UV irradiation at 365 nm resulted in covalent incorporation of [H-3]BP into the nAChR subunits (delta > alpha similar to beta > gamma), 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 6 subunit of delta Phe-232 in delta M1 and delta Pro-286/delta Ile-288 near the beginning of delta M3 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 (alpha Val-255, beta Val-261, and delta Val-269) that was reduced by 90% upon desensitization and labeling in the transmembrane M3 helices of the beta and gamma subunits (beta Met-285, beta Met-288, and gamma Met-291) that was reduced by 50-80% in the desensitized state. Labeling at the lipid interface (alpha Met-415 in alpha M4) 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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据