EFFECTORS OF FUNCTIONAL ORGANIZATION OF TRANSMEMBRANE PR

跨膜 PR 功能组织的影响因素

• 批准号: 3294730
• 负责人: MARINO MARTINEZ-CARRION
• 金额: $ 10.59万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3294730
• 关键词: acetylcholine; bungarotoxins; calorimetry; fluorescent dye /probe; infrared spectrometry; membrane lipids; membrane permeability; membrane reconstitution /synthesis; monoclonal antibody; phospholipids; stop flow technique

Many cationic channels in skeletal and heart muscle are transmembranous proteins. As such they contain structural elements for recognition of the channel opener messenger as well as for modulation of the open and close states for organic and monovalent and divalent cations. Transmembrane proteins contain portions of their structure in a lipid environment and others in media of different ionic strengths and composition inside and outside the cell. Elucidation of the roles of the different sections of the protein molecule is of interest to this project. The proposed work is for 4 basic thrust areas: 1) role of lipids or protein conformation and function, 2) effect of monoclonal antibodies on protein function, 3) identification of structural sites for specific monoclonal antibodies effectors of channels function and 4) study of effects of other protein membrane on states related to channel function. The proposed work will use the acetylcholine receptor as prototype of cationic muscle channel for organic and inorganic cations. In these studies we synthesize and obtain a self consistent image of molecular events for a protein mediated membrane event using the following techniques: 1) physical tools, differential scanning calorimetry, fluorescence dynamics and FT-infrared spectroscopy, 2) kinetic procedures, equilibrium binding, ion flux rates, fluorescence quenching stopped-flow procedures, 3) biological tools, membrane reconstitution, monoclonal antibodies, myoblasts cell culture and membrane fractionation, 4) chemical procedures, peptide fractionation and isolation, chemical characterization of peptides, labeling with affinity ligands, reductive methylation, controlled proteolysis, protein chemistry. These different lines of input are integrated within one single laboratory environment.

骨骼和心肌中的许多阳离子通道都是跨膜的 蛋白质。 因此，它们包含结构性元素以识别 频道开启器Messenger以及开放和关闭的调制 有机和单价和二价阳离子的状态。 跨膜 蛋白质在脂质环境中包含其部分结构的一部分， 在内部具有不同离子优势和构图的媒体中的其他人和 在牢房外。 阐明不同部分的角色 蛋白质分子对该项目感兴趣。 拟议的工作是 对于4个基本推力区域：1）脂质或蛋白质构象的作用和 功能，2）单克隆抗体对蛋白质功能的影响，3） 鉴定特定单克隆抗体的结构位点 通道功能的效应子和4）研究其他蛋白质的作用 与通道功能有关的状态上的膜。 拟议的工作将使用 乙酰胆碱受体作为阳离子肌肉通道的原型 有机和无机阳离子。 在这些研究中，我们合成并获得 蛋白质介导的膜的分子事件的自我一致图像 使用以下技术的事件：1）物理工具，差异 扫描量热法，荧光动力学和FT-Infrared光谱， 2）动力学程序，平衡结合，离子通量速率，荧光 淬火停止流程，3）生物工具，膜 重构，单克隆抗体，成肌细胞培养和膜 分馏，4）化学程序，肽分馏和隔离，， 肽的化学表征，用亲和配体标记， 还原性甲基化，受控蛋白水解，蛋白质化学。 这些 不同的输入线集成在一个单个实验室中 环境。