STRUCTURE OF THE VACUOLAR ATPASE BY ELECTRON MICROSCOPY

描述（改编自摘要）：中性H+-ATP酶代表一种普遍存在的 在亚细胞膜中发现的一类ATP驱动的质子泵 真核细胞的区室。液泡ATP酶的功能是 酸化细胞内部，同时给细胞器的膜提供能量， 例如网格蛋白包被的囊泡、内体、溶酶体、嗜铬颗粒和 高尔基体衍生的小泡。对催化剂的催化机理知之甚少。 这种多亚基复合物，主要是由于缺乏详细的结构， 信息.尽管有各种各样的人类， 像癌症和骨质疏松症这样的基本疾病都与 人空泡型ATP酶活性异常。申请人正在学习 牛脑空泡ATP酶的三维结构 网格蛋白包被囊泡的电子显微镜结合图像 重建方法他的近期目标是：1）获得三分 空泡ATP酶的三维模型，和2）定义的安排 通过使用单克隆抗体标记复合物内的亚基， 相应的多肽。一个长期的目标是将该建筑群的图像 在催化循环的不同阶段，以解决大的 伴随催化转换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.