STRUCTURE/FUNCTION OF ROS PLASMA MEMBRANE PROTEINS

ROS 血浆膜蛋白的结构/功能

• 批准号: 2608555
• 负责人: ROBERT S MOLDAY
• 金额: $ 13.19万
• 依托单位: UNIVERSITY OF BRITISH COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-04-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2608555
• 关键词: SDS polyacrylamide gel electrophoresis; affinity chromatography; autoradiography; calmodulin; chemical binding; chimeric proteins; cone cell; crosslink; cyclic GMP; high performance liquid chromatography; immunocytochemistry; immunoprecipitation; laboratory mouse; laboratory rabbit; membrane structure; membrane transport proteins; protein structure function; retina degeneration; rod cell; visual phototransduction; voltage gated channel; western blottings; yeast two hybrid system

DESCRIPTION (Adapted from applicant's abstract): The outer segment of rod photoreceptor cells consists of a plasma membrane that surrounds a highly organized stack of discs. The long term goal of this research program is to identify and characterize rod outer segment (ROS) plasma membrane proteins and define their role in phototransduction, outer segment structure and stability, metabolic and regulatory and retinal degenerative disease including retinitis pigmentosa. The specific aims are: (1) to study the structure-function relationships of the rod cGMP-gated channel: studies will include (a) analysis of the subunit composition and subunit-subunit interactions of the channel complex; (b) defining the role of the glutamic rich region (GARP) of the beta-subunit in outer segment structure; and (c) elucidating mechanisms that regulate the activity of the channel as part of the phototransduction and adaptational processes; (2) to study the structure, function and regulation of the rod Na+/Ca2+-K+ exchanger: studies will be directed toward determining the role of the large intracellular and extracellular domains of the exchanger in: (a) cation transport function; (b) Ca2+-mediated regulation of exchange activity; and (c) protein-protein interactions that may serve to stabilize the outer segment structure; and (3) to identify and characterize two calmodulin-binding proteins of 67 kDa and 160 kDa in ROS as a first step in determining their role in the Ca2+-dependent regulation of outer segment processes. A variety of current biochemical, molecular biological, cell biological, immunochemical and biophysical methods will be employed along with a unique panel of highly specific monoclonal and polyclonal antibodies and cDNAs to outer segment proteins. The results of these studies should provide new insight into the structure, function and regulation of the cGMP-gated channel, the Na+/Ca2+-K+ exchanger and calmodulin-binding proteins of rod segments and define their role in retinal degenerative diseases.

描述（改编自申请人的摘要）：杆的外段 感光细胞由质膜组成，质膜围绕高度 有组织的光盘堆。 该研究计划的长期目标是 识别和表征视杆外节 (ROS) 质膜蛋白 并定义它们在光转导、外节结构和 稳定性、代谢和调节以及视网膜退行性疾病 包括色素性视网膜炎。 具体目的是：（1）研究 杆状 cGMP 门控通道的结构与功能关系：研究 将包括 (a) 亚基组成和亚基-亚基分析 通道复合体的相互作用； (b) 定义谷氨酸的作用 外节结构中β亚基的丰富区（GARP）；和（三） 阐明调节通道活动的机制 光转导和适应过程； (2) 研究 棒式Na+/Ca2+-K+交换器的结构、功能和调节： 研究将旨在确定大型企业的作用 交换器的胞内和胞外结构域： (a) 阳离子 运输功能； (b) Ca2+介导的交换活动调节；和 (c) 蛋白质-蛋白质相互作用，可能有助于稳定外部 段结构； (3) 识别并表征两个 ROS 中 67 kDa 和 160 kDa 的钙调蛋白结合蛋白作为第一步 确定它们在外节 Ca2+ 依赖性调节中的作用 流程。 目前的各种生化、分子生物学、细胞生物学、 将采用免疫化学和生物物理方法以及独特的 一组高度特异性的单克隆和多克隆抗体以及 cDNA 外段蛋白质。 这些研究结果应该提供新的 深入了解 cGMP 门控的结构、功能和调节 通道、Na+/Ca2+-K+ 交换器和视杆细胞钙调蛋白结合蛋白 细分并定义它们在视网膜退行性疾病中的作用。