PHOSPHODIESTERASE AND GTPASE ACTIVATION IN RETINAL RODS

视网膜杆中磷酸二酯酶和 GTP 酶的激活

• 批准号: 3261556
• 负责人: BERNARD K FUNG
• 金额: $ 18.68万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3261556
• 关键词: alanine transaminase; biological signal transduction; chemical binding; chemical structure function; chromatography; cyclic GMP; genetic manipulation; guanosinetriphosphatases; hybridomas; laboratory mouse; laboratory rabbit; membrane permeability; monoclonal antibody; phosphodiesterases; radioassay; retina degeneration; rhodopsin; rod cell; sodium channel; tissue /cell culture; transducin; vision; visual photoreceptor

The goal of the proposed research is to elucidate the molecular mechanism of visual excitation in vertebrate retinal rods. Phototransduction is now known to involve a light-activated enzume cascade that leads to a rapid reduction of cytosolic cycli GMP and the closure of Na+ channels at the plasma membranes. Central to this process is the activation of three proteins: transducin, cyclic GMP phosphodiesterase, and a postulated cyclic GMP-sensitive channel. The goal of this study is to gain and understanding of the molecular basis of photoexcitation through detailed strucutral and functional analyses of these retinal proteins. We propose to carry out the following studies: (1) Specific monoclonal antibody probes directed against transducin and phosphodiesterase will be used to investigate the structure- function relationships of these proteins, to localize their distributions in photoreceptors, and to study the cause of retinal degenerations. (2) Site-directed chemical probes and anti- synthetic peptdie antibodies will be developed to identify and study the active sites of transducin, and to characterize other cellular GPT-binding homologs. (3) The interaction of the lambda inhibitor and cyclic GMP with phosphodiesterase will be invesigated. The primary structure of the phosphodiesterase subunits will be elucidated by recombinant DNA techniques. (4) Finally, an attempt will be made to isolate and identify the cyclic GMP-sensitive channel. By advancing our knowledge in these four specific areas, our research is expected to contribute to the overall goal of achieving a better understanding of how the interactions of these proteins give rise to activation and termination of the photoresponse in visual cells.

拟议的研究的目的是阐明分子 脊椎动物视网膜杆的视觉激发机理。 现在已知光转录涉及光激活 Enzume Cascade导致胞质Cycli的迅速减少 GMP和Na+通道在质膜上的关闭。 这一过程的核心是三种蛋白质的激活： 透明蛋白，环状GMP磷酸二酯酶和假定的环状 GMP敏感通道。 这项研究的目的是获得和 通过理解光激发的分子基础 这些视网膜的详细诱因和功能分析 蛋白质。 我们建议进行以下研究：（1） 针对转霉素的特异性单克隆抗体探针 磷酸二酯酶将用于研究结构 这些蛋白质的功能关系，以本地化 光感受器中的分布，并研究视网膜的原因 退化。 （2）定向的化学探针和抗 将开发合成PEPTDIE抗体来识别和 研究转霉素的活性位点，并表征其他 细胞GPT结合同源物。 （3）lambda的相互作用 用磷酸二酯酶抑制剂和环状GMP将是 发明。 磷酸二酯酶的主要结构 亚基将通过重组DNA技术阐明。 （4） 最后，将尝试隔离并识别循环 GMP敏感通道。 通过在这四个中促进我们的知识 特定领域，我们的研究有望为 更好地了解如何了解的总体目标 这些蛋白质的相互作用会引起激活和 视觉细胞中光响应的终止。