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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 我们研究了来自眼镜的跨膜水通道Aquaporin 0（AQP0），在突变时会导致镜头，白内障和失明的严重病变。通过与钙调蛋白的相互作用，AQP0通过响应[Ca2+]的变化来调节眼镜内的水渗透性。钙调蛋白（CAM）是一种无处不在的胞质蛋白，通常与CA2+调节结合。我们建议与AQP0的CA2+/CAM相互作用导致水通道内的重大构象变化，使水的渗透率燃烧率。该建议的目的是使用X射线晶体学来确定AQP0-CAM复合物的高分辨率结构。我们可以轻率地种植包含AQP0和CAM的大晶体，但是获得高分辨率的结构信息已被衍射较差所阻碍。在实现我们的目标方面的成功将在水道调节领域具有广泛的影响，以及对CA2+/CAM信号系统的更广泛影响，并促进一般的膜结构生物学。