PERMEABILITY CONTROL OF ACETYLCHOLINE RECEPTOR

乙酰胆碱受体的通透性控制

• 批准号: 6094354
• 负责人: JONATHAN Brewer COHEN
• 金额: $ 5万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-09-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6094354
• 关键词: Torpedo; Xenopus oocyte; affinity labeling; alternatives to animals in research; chemical binding; conformation; electrical measurement; fish electric organ; membrane channels; membrane lipids; membrane permeability; molecular cloning; molecular site; nicotinic receptors; point mutation; protein sequence; protein structure function; radiotracer; receptor binding; site directed mutagenesis; synapses

We wish to understand how the nicotinic acetylcholine receptor (AChR) functions as a ligand-gated ion channel and to understand how drugs and toxins interact with the AChR to alter its function. Torpedo electric tissue will be fractionated to. isolate postsynaptic membranes to be used in biochemical studies designed to: 1) identify functional domains (ligand binding sites, ion channel) in terms of the known amino acid sequences of AChR subunits and to identify differences in structure of those domains that distinguish between the binding of agonists and antagonists or between different conformational states of the AChR, and 2) provide a general description of the three dimensional structure of the AChR in terms of the regions of each subunit exposed at extracellular or cytoplasmic surfaces, at subunit interfaces, or in contact with lipid. Radiolabeled affinity labels and structural probes will be covalently incorporated into AChR, and labeled AChR subunits will be isolated and degraded so that sites of labeling will be determined by N-terminal sequence analysis of isolated labeled peptides. The structural studies will provide a definition of particular amino acids and regions contained within binding sites, at subunit interfaces, or at the protein- lipid interface, but they do not of their own assess the importance of the identified amino acids as determinants of ligand binding affinity or their involvement in the mechanism of channel gating. To address these issues a third research goal is to test models of AChR structure derived from the structural studies by analysis of functional properties of mutant AChRs expressed in Xenopus oocytes. The equilibrium binding affinity of agonists and antagonists will be assessed by radioligand binding assays, while AChR function will be assessed by electrophysiological techniques. Point mutations will be introduced to change amino acids predicted to be important for ligand binding affinity or for channel-gating, and additional mutations will be made of amino acids predicted to be important in the propagation of structural changes from the ACh binding site to the ion channel.

我们希望了解烟碱乙酰胆碱受体（AChR）

项目成果

Interactions between 3-(Trifluoromethyl)-3-(m-[(125)I]iodophenyl)diazirine and tetracaine, phencyclidine, or histrionicotoxin in the Torpedo series nicotinic acetylcholine receptor ion channel.

鱼雷系列烟碱乙酰胆碱受体离子通道中 3-(三氟甲基)-3-(间-[(125)I]碘苯基)二氮丙啶与丁卡因、苯环利定或组烟毒素之间的相互作用。

• DOI: 10.1124/mol.59.6.1514
• 发表时间: 2001
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Gallagher,MJ;Chiara,DC;Cohen,JB
• 通讯作者: Cohen,JB

Mapping the agonist binding site of the nicotinic acetylcholine receptor. Orientation requirements for activation by covalent agonist.

绘制烟碱乙酰胆碱受体的激动剂结合位点。

• DOI: 10.1074/jbc.275.17.12651
• 发表时间: 2000
• 期刊: The Journal of biological chemistry
• 作者: Sullivan,DA;Cohen,JB
• 通讯作者: Cohen,JB

Agonist-induced changes in the structure of the acetylcholine receptor M2 regions revealed by photoincorporation of an uncharged nicotinic noncompetitive antagonist.

不带电荷的烟碱非竞争性拮抗剂的光掺入揭示了激动剂诱导的乙酰胆碱受体 M2 区域结构的变化。

• 发表时间: 1992
• 期刊: The Journal of biological chemistry
• 作者: White,BH;Cohen,JB
• 通讯作者: Cohen,JB

Photoaffinity labeling the torpedo nicotinic acetylcholine receptor with [(3)H]tetracaine, a nondesensitizing noncompetitive antagonist.

用[(3)H]丁卡因（一种非脱敏非竞争性拮抗剂）对鱼雷烟碱乙酰胆碱受体进行光亲和标记。

• DOI: 10.1124/mol.56.2.290
• 发表时间: 1999
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Middleton,RE;Strnad,NP;Cohen,JB
• 通讯作者: Cohen,JB

Conformations of Torpedo acetylcholine receptor associated with ion transport and desensitization.

鱼雷乙酰胆碱受体的构象与离子转运和脱敏相关。

• DOI: 10.1021/bi00257a032
• 发表时间: 1982
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Neubig,RR;Boyd,ND;Cohen,JB
• 通讯作者: Cohen,JB