11 CIS RETINAL ESTER HYDROLASE IN THE EYE

11 眼睛中的顺式视黄醛酯水解酶

• 批准号: 6346183
• 负责人: ANDREW T C TSIN
• 金额: $ 14.87万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6346183
• 关键词: SDS polyacrylamide gel electrophoresis; biosynthesis; chromophore; cis trans isomerization; cow; density gradient ultracentrifugation; enzyme activity; enzyme structure; eye; high performance liquid chromatography; hydrolase; immunoaffinity chromatography; immunocytochemistry; immunofluorescence technique; protein reconstitution; protein sequence; retinal pigment epithelium; retinaldehyde; tissue /cell culture; visual pathways

Background: Current concept of the visual pathway involves the formation of 11-cis retinol from all-trans retinyl esters by an isomerohydrolase. Both 11-cis retinal and 11-cis retinyl esters are derived from this 11-cis retinol and this branch point of the visual pathway is known. Although the fate of 11-cis retinal to serve as visual chromophore has been well studied, exactly how 11-cis retinyl esters are used to supply retinal chromophores for pigment regeneration is not known. Preliminary Studies: Previous studies conducted in our laboratory established the presence of an 11-cis retinyl ester hydrolase (REH) activity in the bovine retinal pigment epithelium (RPE) microsomes. Subcellular fractionation by sucrose and Percol gradients showed that 11- cis retinyl esters and hydrolase enzyme activity are co-localized at the plasma membrane (PM). Hypothesis: These data support the hypothesis that in the RPE PM, there is an 11-cis REH enzyme. This enzyme may control a novel, unexplored branch of the visual pathway to supply visual chromosomes for visual pigment regeneration. Methods: We propose to extend our current research to include complete purification and biochemical characterization of this 11-cis REH enzyme. When a sufficient amount of this protein is available, polyclonal antibody will be raised and immunofluorescence method will be used to study the localization of this enzyme in the RPE. Biomedical Relevance: This research is important because details of this branch of the visual pathway is completely lacking. Results from studies on the control of 11-cis retinyl ester hydrolysis could provide rationale for understanding those ocular diseases associated with a reduction of visual sensitivity and/or delays in the time course of bleaching recovery.

背景：视觉通路的当前概念涉及通过异构水解酶从全反式视黄酯形成 11-顺式视黄醇。 11-顺式视黄醛和11-顺式视黄酯均源自该11-顺式视黄醇，并且视觉通路的该分支点是已知的。尽管 11-顺式视黄醛作为视觉生色团的命运已得到充分研究，但 11-顺式视黄酯如何用于为色素再生提供视网膜生色团的具体情况尚不清楚。初步研究：我们实验室之前进行的研究证实牛视网膜色素上皮 (RPE) 微粒体中存在 11-顺式视黄酯水解酶 (REH) 活性。通过蔗糖和 Percol 梯度进行的亚细胞分级显示，11-顺式视黄酯和水解酶活性共定位于质膜 (PM)。假设：这些数据支持 RPE PM 中存在 11-cis REH 酶的假设。这种酶可能控制视觉通路的一个新的、未经探索的分支，为视色素再生提供视觉染色体。方法：我们建议扩展我们目前的研究，包括这种 11-cis REH 酶的完全纯化和生化表征。当获得足够量的该蛋白时，将产生多克隆抗体，并使用免疫荧光方法研究该酶在 RPE 中的定位。生物医学相关性：这项研究很重要，因为完全缺乏视觉通路这一分支的细节。控制 11-顺式视黄酯水解的研究结果可以为理解那些与视觉敏感性降低和/或漂白恢复时间延迟相关的眼部疾病提供理论基础。