STRUCTURAL CHARACTERIZATION OF CHLORIDE CHANNELS

氯离子通道的结构表征

• 批准号: 3357273
• 负责人: ZVI IOAV CABANTCHIK
• 金额: $ 9.26万
• 依托单位: HEBREW UNIVERSITY OF JERUSALEM
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 1990-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3357273
• 关键词: affinity chromatography; affinity labeling; antibody formation; antiidiotype antibody; biotin; chemical structure function; chemical synthesis; cystic fibrosis; dextrans; enzyme linked immunosorbent assay; haptens; hormone regulation /control mechanism; human tissue; immunochemistry; immunoconjugates; immunofluorescence technique; immunoglobulins; liposomes; macromolecule; membrane activity; membrane permeability; monoclonal antibody; nose neoplasm; o aminobenzoate; oral mucosa; proteolipids; radiotracer; respiratory epithelium; tissue /cell culture; trachea

The present research proposal deals with the biochemical definition of conductive chloride channels in airway epithelia of several animal species and of human nasal tissue. The ultimate goal is to gain understanding of the possible structural changes in the channel components which underlie the deranged activity in epithelia of cystic fibrosis patients. Several basic approaches are proposed to be used for the identification and structural characterization of the putative channel components: (i) affinity labeling based on the use of covalent binding and photoreactive binding analogs of phenyl-anthranilate-related drugs which have demonstrable inhibitory activity on the channels. (ii) immunodetection of the channel components by antiidiotypic antibodies raised against the channel blockers-coupled to carrier proteins. (iii) isolation of the putative components by immunoaffinity chromatography and/or immunoprecipitation of detergent solubilized membranes. (iv) reconstitution of the isolated components into proteoliposomes of defined lipid composition. (v) assessment of the protein activity in the reconstituted state by novel transport techniques of high temporal and detection sensitivity. (vi) assessment of possible modulatory role of ionic and hormonic effectors on the transport function in isolated and reconstituted vesicles in relation to the native function activity. These studies are aimed at providing basic information on chloride channel structure and function and ana experimental model system for evaluating the role played by them in secretory processes of respiratory tissue in both physiological and pathological conditions.

本研究建议涉及生化 气道上皮中的导电氯化物通道的定义 几种动物物种和人类鼻组织。 最终 目标是了解可能的结构变化 基础的通道组件是毁灭性活动的基础 囊性纤维化患者上皮。 建议将几种基本方法用于 假定的识别和结构表征 频道组件： （i）基于共价绑定和 与苯基苯基苯基相关的光反应性结合类似物 对具有明显抑制活性的药物 频道。 （ii）通过抗iDiotiotypic对通道组件进行免疫检测 与载体耦合的通道阻滞剂提出的抗体 蛋白质。 （iii）通过免疫亲和力隔离推定的成分 洗涤剂的色谱和/或免疫沉淀 溶解的膜。 （iv）将孤立的组件重构为 定义的脂质组成的蛋白质体。 （v）评估通过 高时间和检测的新型运输技术 灵敏度。 （vi）评估离子和激素的可能调节作用 隔离和重构的运输功能的效应子 与天然功能活动有关的囊泡。 这些研究旨在提供有关氯化物的基本信息 通道结构和功能以及ANA实验模型 评估他们在分泌中所扮演的角色的系统 生理和 病理状况。