Caveolin in Mammalian Photoreceptor Outer Segements

申请人（申请人）？摘要：总体的长期目标 这项研究的目的是了解 影响光感受器的信号传递。视觉传导调节因子 目前还不完全了解。表征调节蛋白 调节光传导对于理解视网膜和视网膜都至关重要 生理学和病理学。这项研究将研究一个 G蛋白信号传导的潜在重要调节剂，小窝蛋白， 光感受器外节。此应用程序的目标是演示 小窝蛋白是否是真正的外节蛋白，以确定是否 小窝蛋白与感光细胞G蛋白，转导蛋白， 这种关联是否影响转导蛋白活性，并检查， 对小窝蛋白/转导的潜在光/暗介导的体内效应 协会将通过以下方法评估外节中小窝蛋白的存在： 仔细的细胞分离、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来确定， 用纯化的转导素进行直接蛋白质-蛋白质结合实验， 表达caveolin融合蛋白。此外，小窝蛋白衍生的效应 肽对转导素α亚基核苷酸交换和GT活性的影响 将被评估。

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.