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.

骨骼肌和心肌中的许多阳离子通道是跨膜的 proteins. 因此，它们包含了承认 通道开启器信使以及用于调节打开和关闭 状态的有机和单价和二价阳离子。 跨膜 蛋白质在脂质环境中包含其结构的部分， 另一些在不同离子强度和成分的介质中， 在细胞外。 阐明各部门的作用 这个蛋白质分子对这个项目很有意义。 拟议的工作是 对于4个基本的推力领域：1）脂质或蛋白质构象的作用， 功能，2）单克隆抗体对蛋白质功能的影响，3） 特异性单克隆抗体结构位点的鉴定 通道功能的效应物和4）其他蛋白质的作用的研究 与通道功能相关的膜上状态。 拟议的工作将使用 乙酰胆碱受体作为阳离子肌肉通道原型 有机和无机阳离子。 在这些研究中，我们合成并获得了 蛋白质介导膜的分子事件的自洽图像 事件使用以下技术：1）物理工具，差分 扫描量热法、荧光动力学和FT-红外光谱， 2)动力学方法，平衡结合，离子通量，荧光 淬灭停流程序，3）生物工具，膜 重组，单克隆抗体，成肌细胞培养和膜 分级，4）化学方法，肽分级和分离， 肽的化学表征，用亲和配体标记， 还原甲基化，受控蛋白水解，蛋白质化学。 这些 不同的输入线集成在一个实验室中 环境