STRUCTURE OF THE VACUOLAR ATPASE BY ELECTRON MICROSCOPY

电子显微镜观察液泡ATP酶的结构

• 批准号: 6180776
• 负责人: Stephan Wilkens
• 金额: $ 12.26万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6180776
• 关键词: adenosinetriphosphatase; bioimaging /biomedical imaging; cell membrane; conformation; cryoelectron microscopy; electron microscopy; enzyme activity; enzyme complex; enzyme mechanism; enzyme model; enzyme structure; hydrogen transport; hydrogen transporting ATP synthase; physical model; structural biology; vesicle /vacuole

DESCRIPTION (Adapted from abstract): Vacuolar H+-ATPase represents a ubiquitous class of ATP driven proton pumps found in the membrane of subcellular compartments of eukaryotic cells. The function of the vacuolar ATPase is to acidify the interior and at the same time energize the membranes of organelles such as clathrin coated vesicles, endosomes, lysosomes, chromaffin granules and Golgi derived vesicles. Very little is known about the catalytic mechanism of this multi subunit complex, mainly due to the lack of detailed structural information. This deficit exists despite the fact that a wide variety of human diseases as fundamental as cancer and osteoporosis are associated with an abnormal activity of the human vacuolar type ATPase. The applicant is studying the three-dimensional structure of the vacuolar ATPase from bovine brain clathrin-coated vesicles by electron microscopy in conjunction with image reconstruction methods. His immediate goals are: 1) to obtain a three dimensional model of the vacuolar ATPase, and 2) to define the arrangement of the subunits within the complex by using monoclonal antibodies to tag the corresponding polypeptides. A longer-term objective is to image the complex at different stages in the catalytic cycle in order to resolve the large conformational changes that accompany catalytic turnover and energy coupling.

描述（根据摘要改编）：液泡H+-ATPase代表无处不在的 亚细胞膜中发现的ATP驱动质子泵 真核细胞的隔室。液泡ATPase的功能是 内部和同时酸化细胞器的膜 例如网格蛋白涂层的囊泡，内体，溶酶体，铬蛋白颗粒和 高尔基衍生的囊泡。关于催化机理的了解很少 这个多亚基复合物，主要是由于缺乏详细的结构 信息。尽管有各种各样的人类，但存在这种赤字 与癌症和骨质疏松症一样基本的疾病与 人液泡型ATPase的异常活性。申请人正在学习 来自牛脑的液泡ATPase的三维结构 通过电子显微镜与图像结合的网状蛋白涂层囊泡 重建方法。他的直接目标是：1）获得三个 液泡ATPase的尺寸模型，2）定义排列 通过使用单克隆抗体来标记复合物中的亚基 相应的多肽。一个长期目标是对复合物进行图像 催化周期中的不同阶段以解决大型 催化转换和能量耦合伴随的构象变化。