Molecular Basis for Rod Outer Segment Structure and Retinal Degenerative Diseases

视杆外节结构和视网膜退行性疾病的分子基础

基本信息

批准号：
7633188
负责人：
ROBERT S MOLDAY
金额：
$ 26.85万
依托单位：
UNIVERSITY OF BRITISH COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-04-01 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7633188
关键词：
Adhesions Affinity Anabolism Area Binding Biochemical Biological Assay Blindness C-terminal Cataloging Catalogs Cell Adhesion Cell Adhesion Molecules Cell Fractionation Cell Survival Cell membrane Cells Cellular biology Complex Cyclic GMP Defect Degenerative Disorder Development Disease Elements Enzymes Event Extracellular Matrix Filament Genes Glutamic Acid Goals Inherited Intervention Knockout Mice Light Adaptations Link Macular degeneration Mass Spectrum Analysis Mediating Membrane Metabolism Molecular Morphogenesis Mutation Photoreceptors Phototransduction Play Procedures Process Production Property Protein Biosynthesis Proteins Proteomics Research Retinal Retinal Cone Retinal Degeneration Retinal Diseases Retinal Dystrophy Retinal Photoreceptors Retinitis Pigmentosa Retinoids Rod Outer Segments Role Signal Transduction Site-Directed Mutagenesis Stargardt&apos s disease Structure Synapses Techniques Triton Vertebrate Photoreceptors Very Long Chain Fatty Acid Vision X-Linked Retinoschisis XLRS1 protein base design insight lipid biosynthesis macular dystrophy membrane reconstitution multicatalytic endopeptidase complex mutant novel numb protein outer plexiform layer peripherin photoreceptor degeneration potassium exchanger sodium-calcium programs protein function protein protein interaction retinal neuron retinal rods synaptic function trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): Outer segments are specialized compartments of rod and cone photoreceptors that mediate the primary events of vision. The rod outer segment (ROS) consists of a highly ordered stack of disks surrounded by a plasma membrane. Filaments bridge adjacent disks together and the disk rim to the plasma membrane, however, the proteins that constitute these structural elements have not been identified. Since disorganization and loss in ROS is known to cause photoreceptor degeneration, it is important to define the molecular basis for ROS morphogenesis and structure. An overall objective of this research is to catalogue all the proteins in ROS with the goal of identifying a subset of "novel" proteins critical for the structure, stabilization and formation of ROS and proteins likely to be associated to retinal degenerative diseases. Another key objective is to define the molecular basis for selected inherited retinal degenerative diseases that cause significant loss in vision. These objectives will be achieved in the following specific aims. 1) To determine the complete proteosome of the ROS using current and emerging mass spectrometry-based approaches together with subcellular fractionation procedures. Proteins predicted to play a role in ROS structure and morphogenesis will be characterized using an array of biochemical, immunochemical, molecular and cell biology techniques. The role of these proteins in ROS structure and photoreceptor degeneration will be assessed; 2) To evaluate the role of protein-protein interactions between the peripherin-2:rom-1 complex in the disk rim and the channel:exchanger complex in the plasma membrane in stabilizing the structure of ROS. Immuno-affinity techniques, site-directed mutagenesis, heterologous cell expression, membrane reconstitution and knockout mice will be used in the study; and 3) to examine the molecular mechanisms underlying selected retinal diseases. The function of RS1 (retinoschisin) as a retinal cell adhesion protein will be examined in order to understand how defects in this protein cause X-linked Juvenile Retinoschisis. The possible function of ELOVL4 in the biosynthesis of very long chain fatty acids and in autosomal dominant Stargardt disease will also be studied. This research will provide new insight into the molecular mechanisms underlying ROS structure and inherited retinal dystrophies and serve as a basis for developing novel treatments for this set of diseases that cause significant visual loss.

描述（由申请人提供）：外节是介导视觉主要事件的杆状和锥状光感受器的特殊隔室。杆外节（ROS）由高度有序的圆盘堆叠组成，周围有质膜。细丝将相邻的圆盘桥接在一起并将圆盘边缘连接到质膜，然而，构成这些结构元件的蛋白质尚未被鉴定。由于已知 ROS 的解体和损失会导致光感受器变性，因此定义 ROS 形态发生和结构的分子基础非常重要。这项研究的总体目标是对 ROS 中的所有蛋白质进行编目，目的是确定对 ROS 的结构、稳定和形成至关重要的“新”蛋白质子集以及可能与视网膜退行性疾病相关的蛋白质。另一个关键目标是确定导致视力严重丧失的选定遗传性视网膜退行性疾病的分子基础。这些目标将通过以下具体目标来实现。 1) 使用当前和新兴的基于质谱的方法以及亚细胞分级分离程序来确定 ROS 的完整蛋白体。预计在 ROS 结构和形态发生中发挥作用的蛋白质将使用一系列生物化学、免疫化学、分子和细胞生物学技术进行表征。将评估这些蛋白质在 ROS 结构和光感受器变性中的作用； 2) 评估盘缘中的外周蛋白-2:rom-1复合物与质膜中的通道:交换器复合物之间的蛋白质-蛋白质相互作用在稳定ROS结构中的作用。研究中将使用免疫亲和技术、定点突变、异源细胞表达、膜重建和基因敲除小鼠； 3）检查选定视网膜疾病的分子机制。我们将检查 RS1（视网膜劈裂素）作为视网膜细胞粘附蛋白的功能，以了解该蛋白质的缺陷如何导致 X 连锁青少年视网膜劈裂症。还将研究 ELOVL4 在极长链脂肪酸生物合成和常染色体显性 Stargardt 病中的可能功能。这项研究将为 ROS 结构和遗传性视网膜营养不良的分子机制提供新的见解，并为开发针对这一系列导致严重视力丧失的疾病的新疗法奠定基础。