Caveolin in Mammalian Photoreceptor Outer Segements

简介(摘自申请人？S摘要)：总体目标、长远目标 这项研究的目的是了解其作用机制和途径。 影响光感受器中的信号。视觉传导调节因子 目前还不能完全理解。调控蛋白的特性研究 调制光信号转导对于理解视网膜和视网膜 生理学和病理学。这项拟议的研究将考察 潜在重要的G蛋白信号调节剂，小窝蛋白，在 光感受器外节。本应用程序的目标是演示 小窝蛋白是否是一个可靠的外段蛋白，以确定 小窝蛋白与光感受器G蛋白，转导蛋白相关联，以确定 这种联系是否影响转导蛋白的活性，并检查 在体内，潜在的光/暗对小窝蛋白/转导的影响 协会。将通过以下方式评估外侧节段中是否存在凹陷 仔细进行细胞分级、免疫印迹分析和免疫细胞化学。 小窝蛋白/转导蛋白的相互作用将通过免疫沉淀和 纯化的转导蛋白与蛋白质的直接结合实验 表达小窝蛋白融合蛋白。此外，洞穴土衍生的效果 多肽对转导蛋白α亚基核苷酸交换和GTP酶活性的影响 将会被评估。

DESCRIPTION (From the Applicant?s Abstract): The overall, long-term objective of this research is to gain an understanding of the mechanisms and pathways that influence signaling in photoreceptors. Regulators of visual transduction are incompletely understood at present. Characterizing the regulatory proteins that modulate phototransduction is essential in understanding both retinal physiology and pathology. The research proposed will examine the influence of a potentially important modulator of G-protein signaling, caveolin, in photoreceptor outer segments. The goals of this application are to demonstrate whether caveolin is a bon fide outer segment protein, to determine whether caveolin associates with the photoreceptor G-protein, transducin, to determine whether this association influences transducin activity, and to examine, in vivo, potential light/dark-mediated effects on caveolin/transducing association. The presence of caveolin in outer segments will be assessed by careful cell fractionation, immunoblot analysis ad immunocytochemistry. Caveolin/transducin interactions will be determined by immunoprecipitation and direct protein-protein binding experiments with purified transducin and expressed caveolin fusion protein. In addition, the effect of caveolin-derived peptides on nucleotide exchange and GTPase ctivity of transducin alpha subunits will be assessed.