CRYSTALLOGRAPHIC STUDIES OF THE AQP0-CAM COMPLEX

AQP0-CAM 复合体的晶体学研究

• 批准号: 7954443
• 负责人: Stephen Loen Reichow
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954443
• 关键词: Blindness; Calmodulin; Cataract; Complex; Computer Retrieval of Information on Scientific Projects Database; Coupled; Crystalline Lens; Funding; Grant; Institution; Lead; Lesion; MIP gene; Membrane; Mutate; Permeability; Proteins; Regulation; Research; Research Personnel; Resolution; Resources; Signal Transduction; Source; Structure; System; United States National Institutes of Health; Water; X-Ray Crystallography; lens; structural biology; success; synchrotron radiation; water channel

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们研究水通道蛋白 0 (AQP0)，这是一种来自眼睛晶状体的跨膜水通道，突变时会导致晶状体严重损伤、白内障和失明。 AQP0 通过与钙调蛋白相互作用来响应 [Ca2+] 的变化，从而调节眼晶状体内的水渗透性。钙调蛋白 (CaM) 是一种普遍存在的胞质蛋白，通常与 Ca2+ 调节相关。我们认为 Ca2+/CaM 与 AQP0 的相互作用导致水通道内的主要构象变化，从而影响水的渗透率。该提案的目的是利用 X 射线晶体学来确定 AQP0-CaM 复合物的高分辨率结构。我们可以完全生长含有 AQP0 和 CaM 的大晶体，但是由于衍射较差，无法获得高分辨率的结构信息。成功实现我们的目标将对水通道调节领域产生广泛影响，并对 Ca2+/CaM 信号系统和整体膜结构生物学产生更广泛的影响。