MOLECULAR CHARACTERIZATION OF CHLORIDE CHANNEL PROTEINS

该项目的总体目标是从分子水平上了解 氯离子通道与健康和疾病。氯离子通道起着至关重要的作用 在许多细胞和组织过程中发挥作用；氯通道功能障碍 已知在人类疾病中具有主要致病意义 囊性纤维化、先天性肌强直和霍乱。充分了解 氯离子通道的结构和功能将提供对 研究这些疾病的发病机制，并可能提出新的治疗方法。 我们克隆了编码kD氯离子通道蛋白的基因。 牛肾。该基因的表达是表达 卵泡刺激物诱导非洲爪哇卵母细胞cAMP激活的氯通道 纤维化跨膜电导调节剂(CFTR)。这样做的目的是 建议研究P64及其在各种氯离子通道中的作用 包括与cftr相关的cAMP激活的氯通道。这个 具体目的是：1)表征与P64相关的氯离子通道 通过免疫沉淀、重组和体内磷酸化，2) 分离编码P64的人类同源物的cDNA，以及3)推导出 利用反义抑制P64基因表达的表达系统 内源基因与野生型和突变型P64的重新导入 旨在逃避反义抑制。

The overall goal of the project is to gain a molecular understanding of chloride channels in health and disease. Chloride channels play essential roles in many cell and tissue processes; chloride channel dysfunction is known to be of primary pathogenic significance in the human diseases cystic fibrosis, myotonia congenita, and cholera. A full understanding of chloride channel structure and function will provide insight into the pathogenesis of these diseases and may suggest new therapeutic approaches. We have cloned the cDNA encoding a 64 kD chloride channel protein from bovine kidney. Expression of this gene is required for expression of the cAMP-activated chloride channel induced in Xenopus oocytes by the cystic fibrosis transmembrane conductance regulator (CFTR). The purpose of this proposal is to study P64 and its role in various chloride channels including the cAMP-activated chloride channel associated with CFTR. The specific aims are to: 1) Characterize the P64 associated chloride channel by immunoprecipitation, reconstitution and in vivo phosphorylation, 2) Isolate cDNAs encoding the human homologue of P64, and 3) Derive an expression system for P64 using antisense suppression of expression of the endogenous gene and reintroduction of wild type or mutated versions of P64 designed to evade the antisense suppression.