CRYSTALLOGRAPHIC STUDIES OF THE AQP0-CAM COMPLEX

AQP0-CAM 复合体的晶体学研究

• 批准号: 8362162
• 负责人: Stephen Loen Reichow
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362162
• 关键词: Blindness; Calmodulin; Cataract; Complex; Coupled; Crystalline Lens; Funding; Grant; Lead; Lesion; MIP gene; Membrane; Mutate; National Center for Research Resources; Permeability; Principal Investigator; Proteins; Radiation; Regulation; Research; Research Infrastructure; Resolution; Resources; Signal Transduction; Source; Structure; System; United States National Institutes of Health; Water; X-Ray Crystallography; cost; lens; structural biology; success; water channel

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We study aquaporin 0 (AQP0), a transmembrane water channel from the eye lens, causing severe lesions in the lens, cataracts and blindness when mutated. AQP0 regulates water permeability within the eye lens by responding to changes in [Ca2+] through an interaction with calmodulin. Calmodulin (CaM) is a ubiquitous cytosolic protein commonly coupled with Ca2+ regulation. We suggest the Ca2+/CaM interaction with AQP0 lead to major conformational changes within the water channels that enfluence water permeability rates. The aims of this proposal are to use X-ray crystallography to determine the high-resolution structure of the AQP0-CaM complex. We can fatihfully grow large crystals containing both AQP0 and CaM, however obtaining high-resolution structural information has been stymied by poor diffraction. Success in achieving our aim will have wide implications in the fields of water channel regulation, as well as broader implications to the Ca2+/CaM signaling system and to furthering membrane structural biology in general.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们研究水通道蛋白0（AQP 0），一种来自眼睛透镜的跨膜水通道，当突变时会导致透镜严重病变、白内障和失明。AQP 0通过与钙调蛋白相互作用响应[Ca 2 +]的变化来调节眼透镜内的水渗透性。钙调素（CaM）是一种普遍存在的细胞质蛋白，通常与Ca 2+调节偶联。我们认为Ca 2 +/CaM与AQP 0的相互作用导致水通道内的主要构象变化，从而影响水渗透率。该提案的目的是使用X射线晶体学来确定AQP 0-CaM复合物的高分辨率结构。我们可以疲劳地生长含有AQP 0和CaM的大晶体，然而，获得高分辨率的结构信息一直受到衍射不良的阻碍。成功实现我们的目标将在水通道调节领域产生广泛的影响，以及对Ca 2 +/CaM信号系统和进一步促进膜结构生物学的广泛影响。