Biosynthesis and trafficking of phosphodiesterase in the retinal photoreceptors

项目概要/摘要 我们研究的长期目标是研究翻译后蛋白质修饰的作用及其对蛋白质表达的影响。 在光传导蛋白质合成和成熟中的作用。本提案的目的是调查 磷酸二酯酶-6（PDE 6）异戊二烯化在视网膜感光神经元中的重要性。PDE 6是 一种对视力至关重要的酶，它的缺失或突变会导致人类视网膜色素变性 (RP)和色盲。有趣的是，芳香烃受体蛋白样-1（AIPL 1），一种共 PDE 6的伴侣蛋白导致更严重的儿童失明。尽管与几起致盲事件有明显联系 疾病，我们对感光神经元中这种必需酶的组装和成熟的知识是 不知道。我们的研究将特别关注AIPL 1，HSP 90和脂质修饰之间的相互作用， PDE 6组装、成熟和运输至光感受器外节。我们亦会研究 差异异戊二烯化在光感受器细胞中介导光依赖性信号传导的重要性。 我们将使用独特的动物模型，体内电生理测量，离体 视网膜培养模型和体外生化分析，以全面解决这个问题。 我们提出的研究与NEI的视网膜疾病计划相一致，以“确定基因 涉及遗传性和视网膜退行性疾病（包括RP），确定 这些突变的病理生理机制，并确定新的潜在的治疗 治疗策略，如基因转移，组织和细胞移植，生长因子治疗， 药物干预”。我们提出的研究具有临床意义，例如治疗 PDE 6和AIPL 1的突变导致人类失明。

PROJECT SUMMARY/ABSTRACT The long term goals of our research are to study the role of post-translational protein modifications and their role in phototransduction protein synthesis and maturation. The purpose of this proposal is to investigate the importance of the isoprenylation of phosphodiesterase-6 (PDE6) in retinal photoreceptor neurons. PDE6 is an enzyme that is essential for vision, and its absence or mutations in humans lead to retinitis pigmentosa (RP) and color blindness. Interestingly, changes in Aryl hydrocarbon receptor protein-like-1 (AIPL1), a co- chaperone for PDE6 leads to more severe childhood blindness. Despite the clear link to several blinding diseases, our knowledge on assembly and maturation of this essential enzyme in photoreceptor neurons is not known. Our study will specifically focus on the interplay between AIPL1, HSP90, and lipid modification in PDE6 assembly, maturation, and trafficking to the photoreceptor outer segments. We will also study the importance of differential isoprenylation in mediating light-dependent signaling in the photoreceptor cells. We will use a combination of unique animal models, in vivo electrophysiological measurements, ex vivo retinal culture models and in vitro biochemical analyses to comprehensively address this question. Our proposed studies are aligned with the Retinal Diseases Program of the NEI to “Identify the genes involved in both inherited and retinal degenerative diseases (including RP), determine the pathophysiological mechanisms underlying these mutations, and determine new potential therapeutic strategies for treatment such as gene transfer, tissue and cell transplantation, growth factor therapy, and pharmacological intervention.” Our proposed studies have clinical implications, such as therapy for mutations in PDE6 and AIPL1 that lead to blindness in humans.