THE ROLE OF AQUAPORIN-0 IN LENS DEVELOPMENT AND AGING

Aquaporin-0 在晶状体发育和老化中的作用

基本信息

批准号：
7645695
负责人：
Kevin L Schey
金额：
$ 30.97万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-05-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7645695
关键词：
Adhesions Affect Age Aging Aging-Related Process Arts Binding C-terminal Calmodulin Cataract Cell Adhesion Cell Adhesion Molecules Cell Differentiation process Confocal Microscopy Cytoskeletal Proteins Development Elements Fluorescence Functional disorder Genes Goals Homeostasis Human Indium Integral Membrane Protein Knock-out Knockout Mice Knowledge Laboratories Lead Lens Fiber MIP gene Maintenance Maps Measures Membrane Membrane Proteins Microscopy Modification Molecular Mus Mutation Peptides Permeability Phosphorylation Play Post-Translational Protein Processing Property Proteins Proteome Proteomics Reporting Research Research Personnel Resolution Role Site Solubility Spatial Distribution Spectrum Analysis Staging Structure Surface Plasmon Resonance Testing Tissues Water age related congenital cataract crosslink deamidation fiber cell filensin improved in vivo laser capture microdissection lens lens transparency prevent programs protein function protein protein interaction

项目摘要

DESCRIPTION (provided by applicant): The human lens must maintain transparency over many decades and, in order to do so, fiber cells must establish close packing and preserve protein solubility. Lens membrane proteins, such as transporters and adhesion molecules, play essential roles in lens development and maintenance of homeostasis. Aquaporin-0 (AQPO) and MP20, the most abundant lens membrane proteins, have reported roles in fiber cell adhesion, in water permeability, and in gap junctional organization and, as such, play important roles in the development and maintenance of lens transparency. The long-term goal of our research is to identify modifications to the lens membrane proteome during development and aging and to understand how lens membrane protein function is altered by modification. Our general hypothesis is that modifications to AQPO and MP20 alter protein function in specific lens regions during lens development and accumulate with age leading to cataracts. More specifically, we hypothesize that AQPO C-terminal modifications regulate calmodulin binding and alter a newly discovered interaction withlens specific cytoskeletal elements in specific regions of the lens. To test this hypothesis our goal is to use state-of-the-art proteomics and microscopy approaches to generate a molecular level understanding of how AQPO and MP20 are modified in distinct stages of fiber cell development. We will then determine how these modifications affect AQPO protein-protein interactions. We propose four aims: 1) To identify AQPO modifications in normal and cataractous human lenses with high spatial resolution corresponding to different stages of fiber cell differentiation and age, and 2) To determine the sites of interaction between AQPO and filensin/CP49, the effect of AQPO modification on this interaction, and the in vivo effects of loss of this interaction, 3) To determine the effects of AQPO modification on interactions with calmodulin, and 4) To identify MP20 modifications in normal and cataractous human lenses with high spatial resolution corresponding to different stages of fiber cell differentiation and age. The global approach proposed is expected to provide new detailed information on the structure and function of the most abundant integral membrane proteins in the lens and lead to an improved understanding of normal lens development, aging processes, and cataractogenesis.

描述（由申请人提供）：人镜必须在数十年中保持透明度，为此，纤维细胞必须建立近距离包装并保留蛋白质溶解度。晶状体膜蛋白（例如转运蛋白和粘附分子）在镜头发育和维持体内平衡中起着至关重要的作用。水通道蛋白-0（AQPO）和MP20是最丰富的透镜膜蛋白，报告了在纤维细胞粘附，水的渗透性以及间隙连接结构组织中的作用，因此在透镜透明度的开发和维持中起着重要作用。我们研究的长期目标是在发育和衰老过程中确定对晶状体膜蛋白质组的修饰，并了解如何通过修饰改变晶状体膜蛋白功能。我们的一般假设是，对AQPO和MP20的修改会在特定晶状体区域的发育过程中改变蛋白质功能，并随着年龄的形式积累而导致白内障。更具体地说，我们假设AQPO C末端修饰调节钙调蛋白结合并改变了与镜头特定区域中特定细胞骨架元素的新发现的相互作用。为了检验这一假设，我们的目标是使用最先进的蛋白质组学和显微镜方法来产生分子水平的理解，以了解AQPO和MP20如何在纤维细胞开发的不同阶段进行修饰。然后，我们将确定这些修饰如何影响AQPO蛋白 - 蛋白质相互作用。我们提出四个目标：1）确定具有高空间分辨率的正常和白内障人类镜头的AQPO修饰，与纤维细胞分化和年龄的不同阶段相对应，以及2）确定AQPO和FILENSIN/CP49之间的相互作用位点，以及AQPO修改对这种交互作用的影响，并确定vivo效应的影响，并确定vivo的损失。与钙调蛋白的相互作用，以及4）在正常和白内障人透镜中识别具有高空间分辨率的MP20修饰，对应于纤维细胞分化和年龄的不同阶段。预计所提出的全球方法将提供有关晶状体中最丰富的整体膜蛋白的结构和功能的新详细信息，并导致人们对正常晶状体发展，衰老过程和白内障生成的理解有了改善。