MOLECULAR BIOLOGY OF CL CHANNEL IN CYSTIC FIBROSIS

囊性纤维化中 CL 通道的分子生物学

• 批准号: 3105885
• 负责人: QAIS AL-AWQATI
• 金额: $ 65.95万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-30 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3105885
• 关键词: chloride channels; chlorine; cystic fibrosis; molecular pathology

Cystic fibrosis is now known to be due to a recessive genetic defect in the regulation of anion transport in epithelial cells. In these epithelia, chloride is transported across the cell membrane through a channel that can be opened by cyclic AMP dependent protein kinase. Recently, it was shown that in cystic fibrosis, the kinase fails to open the channel, even though the channel is present in the membrane. The Columbia-Presbyterian Cystic Fibrosis Center is composed of a multidisciplinary group of investigators who are collaborating towards identification of the molecular defect in this disease. We have already purified the epithelial chloride channel and using this procedure we will clone and sequence the gene for the channel. Methods have been developed for expression of ion transport proteins in frog oocytes which will facilitate the cloning of the chloride channel. Another expression system was developed which relies on DNA-mediated gene transfer, that will allow the cloning of iodide and chloride transport proteins from thyroid epithelial cells. This method will also be used to identify structural and regulatory mutants of these genes. Regulation of the chloride channel by a variety of protein kineses will be studied in human epithelial cells and in cells derived from patients with cystic fibrosis. The chloride channel will be reconstituted in planar lipid bilayer and the effect of kineses on its electrophysiological characteristics will be studied. The Center will promote the collaboration of all the involved groups towards the identification of the genetic defect in cystic fibrosis.

囊性纤维化现在已知是由于隐性遗传 上皮细胞阴离子转运调节缺陷。 在这些上皮细胞中，氯通过细胞转运 通过环腺苷酸可以打开的通道 依赖性蛋白激酶 最近，研究表明，在囊性病变中， 纤维化时，激酶不能打开通道，即使 通道存在于膜中。 哥伦比亚长老会囊性纤维化中心是由一个 一个多学科的研究小组， 来鉴定这种疾病的分子缺陷。 我们已经纯化了上皮氯离子通道， 在这个过程中，我们将克隆和测序通道的基因。 已经开发了表达离子转运的方法， 青蛙卵母细胞中的蛋白质，这将有助于克隆 氯离子通道 开发了另一种表达系统， 依赖于DNA介导的基因转移， 甲状腺上皮细胞碘和氯转运蛋白 细胞 该方法还将用于识别结构和 这些基因的调控突变体。 氯化物的调节 将在人类中研究各种蛋白质激酶对通道的影响 上皮细胞和来源于囊性癌患者的细胞中 纤维化 氯离子通道将在平面内重建， 脂双层膜及其运动对电生理的影响 将研究其特点。 该中心将促进所有相关人员的合作 基因缺陷的鉴定。 纤维化