BIOCHEMISTRY OF THE RETINA & PIGMENT EPITHELIUM

视网膜的生物化学

• 批准号: 3256697
• 负责人: JOHN C SAARI
• 金额: $ 23.83万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-04-01 至 1990-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3256697
• 关键词: biological transport; chromatography; dark adaptation; enzyme mechanism; histochemistry /cytochemistry; immunochemistry; laboratory rabbit; laboratory rat; ligands; mixed tissue /cell culture; nutrition related tag; protein sequence; radioassay; retina; retinal pigment epithelium; retinaldehyde; retinoate; retinoid binding proteins; tissue /cell culture; ultracentrifugation; visual photoreceptor; visual photosensitivity; vitamin A deficiency; vitamin metabolism

The overall goal of this research project is to provide an understanding of the molecular details and control features of the reactions of vitamin A known as the visual cycle. The major emphasis of the work is associated with the hypothesis that vitamin A transport and metabolism is facilitated by a group of retinoid-binding proteins which are postulated to solubilize and carry the vitamin during inter- and intracellular transport and to be involved in the control of its enzymatic transformation. Each of the specific aims pursues an aspect of this central theme. Independent confirmation of a role for Muller cells in the metabolism of vitamin A will be sought employing Muller cells in culture or isolated from suspension of disaggregated retinal cells. Antibodies specific for four different retinoid-binding proteins (CRBP, cellular retinol-binding protein; CRABP, cellular retinoic acid-binding protein; CRALBP, cellular retinaldehyde-binding protein; IRBP, interphotoreceptor retinoid-binding protein) will be used to assess whether a variation in their level occurs following a physiologic (bleaching), nutritional (vitamin A deficiency) or pathologic challenge. The endogenous retinoids associated with three of the retinoid-binding proteins (CRBP, CRALBP and IRBP) will be examined from dark-adapted and bleached cattle eyes or eyes from laboratory animals. A study of the role of retinoid-binding proteins as substrate carriers for enzymatic reactions will be continued with emphasis placed on attempts to demonstrate the specificity of interaction between binding protein and enzyme and the linking of two or more enzymatic reactions. Amino acid sequence studies will be carried out on CRALBP to determine if it is a member of the family of structurally related proteins which now includes CRABP, CRBP, Z-protein and myelin P2 protein. 125I-SRBP (serum retinol-binding protein) will be labeled with a photosensitive ligand and used to probe the molecular nature of the SRBP receptor reported to be present on the basal surface of retinal pigment epithelium.

本研究项目的总体目标是提供对 维生素A反应的分子细节及其调控特征 被称为视觉循环。这项工作的主要重点是与 假设维生素A的运输和新陈代谢促进 通过一组维甲酸结合蛋白，它们被认为可以溶解 并在细胞间和细胞内运输过程中携带维生素 参与了其酶促转化的控制。每一位 《特定目标》追求这一中心主题的一个方面。 米勒细胞在新陈代谢中作用的独立确认 维生素A将通过在培养中使用穆勒细胞或从 悬浮分解的视网膜细胞。四种特异性抗体 不同的视黄醇结合蛋白(CRBP、细胞视黄醇结合 蛋白质；CRABP，细胞维甲酸结合蛋白；CRALBP，细胞 视黄醛结合蛋白；IRBP，光感受器间类视黄酸结合 蛋白质)将被用来评估其水平是否发生变化 生理性(漂白)、营养性(维生素A缺乏)或 病理性挑战。内源性维甲酸与三种 维甲酸结合蛋白(CRBP、CRALBP和IRBP)将从 暗适应和漂白的牛眼或实验室动物的眼睛。一个 视黄酸结合蛋白作为底物载体的作用研究 酶反应将继续进行，重点是尝试 论证结合蛋白与蛋白质相互作用的特异性 酶和两个或多个酶反应的连接。氨基酸 将对CRALBP进行序列研究，以确定它是否是一种 结构上相关的蛋白质家族的成员，现在包括 CRABP、CRBP、Z蛋白和髓鞘P2蛋白。125I-SRBP(血清 视黄醇结合蛋白)将被光敏配体标记并 用于探测SRBP受体的分子性质，据报道 存在于视网膜色素上皮的基面。