3D ELECTRON MICROSCOPY OF THE ATPase HRS-2

ATPase HRS-2 的 3D 电子显微镜

• 批准号: 6622949
• 负责人: PAWEL A. PENCZEK
• 金额: $ 28.25万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6622949
• 关键词: adenosine triphosphate; adenosinetriphosphatase; bioimaging /biomedical imaging; chemical association; conformation; cryoelectron microscopy; electron microscopy; enzyme activity; enzyme inhibitors; monomer; mutant; phosphoproteins; point mutation; protein binding; protein protein interaction; protein structure function; structural biology; syntaxin

DESCRIPTION (provided by applicant): Hrs-2 is an ATPase that has been implicated in a variety of cellular processes by virtue of its functional protein interactions. We have determined that the form of hrs-2 with the highest specific ATPase activity is an oligomer of six subunits. However, the structure of this complex is unknown. Consequently, the extensive functional studies lack a structural basis for their interpretation. Our goal is to use three-dimensional electron microscopy to determine the structure of hrs-2 and its functional complexes. We show in the preliminary results that hrs-2 can be isolated in sufficient quantities to visualize it in the electron microscope and that the data collected is of sufficient quality to perform a three-dimensional reconstruction of this protein. Association of hrs-2 with interacting proteins as well as its intrinsic ATPase activity likely plays a role in its function. Within the framework of this proposal, we will investigate the role of the ATPase activity and interacting proteins in the structure of the hrs-2 oligomer. To achieve this goal, we will employ three-dimensional electron microscopy and further refine the results using computational structure prediction methods. This structural information will provide insight into the role of interacting proteins and enzyme activity on the conformation of the hrs-2 oligomer and ultimately on its function. The Specific Aims are: (1) Three-dimensional electron microscopy of the native structure of hrs-2. (2) High-resolution structure of the hrs-2 by cryo-electron microscopy and structure prediction methods. (3) Determination of the role of hrs-2-interacting proteins on the structure of hrs-2. (4) Determination of the effect of ATP binding on the structure of hrs-2. (5) Determination of the structural basis for the mechanism by which hrs-2 inhibits SNARE complex formation.

描述（由申请人提供）：Hrs-2是一种atp酶