STRUCTURE OF THE VACUOLAR ATPASE BY ELECTRON MICROSCOPY

描述(摘自摘要)：液泡H+-ATPase代表一种无处不在的 亚细胞膜中发现的一类三磷酸腺苷驱动的质子泵 真核细胞的隔间。液泡型ATPase的功能是 使细胞器内部酸化，同时为细胞器的膜提供能量 如包被网状蛋白的囊泡、内小体、溶酶体、嗜铬粒和 高尔基体衍生出小泡。对其催化机理知之甚少。 这种多亚基复合体，主要是由于缺乏详细的结构 信息。这种缺陷是存在的，尽管有各种各样的人类 像癌症和骨质疏松症这样的基础疾病与 人类空泡型ATPase活性异常。申请者正在学习 牛脑空泡ATPase的三维结构 电子显微镜结合图像观察笼状蛋白包裹的囊泡 重建方法。他近期的目标是：1)拿到3分 液泡ATPase的空间模型，以及2)定义 通过使用单抗标记复合体中的亚基 相应的多肽。一个较长期的目标是在 在催化循环的不同阶段，以解决大的 伴随着催化周转和能量耦合的构象变化。

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.