STRUCTURAL ANALYSIS OF THE DYNEIN ATPASE

动力蛋白ATP酶的结构分析

• 批准号: 3280581
• 负责人: KENNETH ALLEN JOHNSON
• 金额: $ 18.16万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280581
• 关键词: Tetrahymena; X ray crystallography; adenosinetriphosphatase; binding proteins; cilium /flagellum motility; cytoskeleton; dynein ATPase; electron microscopy; enzyme linked immunosorbent assay; enzyme structure; gel electrophoresis; intracellular transport; ion exchange chromatography; laboratory mouse; monoclonal antibody; protein structure; proteolysis; scanning electron microscopy

The overall goals of the proposed research are to establish the structure and mechanism of action of the dynein ATPase in generating a force for ciliary and flagellar movement. The specific goals of these studies can be divided into five parts: (1) Localize the polypeptides, the ATPase sites, and the microtubule-binding sites in terms of the three-headed bouquet of the 22S dynein; (2) evaluate the flexibility of the strands connecting the dynein heads to the base; (3) determine the arrangement of the dynein heads on the microtubule surface lattice and the nature of the interaction in the rigor complex; (4) examine the ATP-dependent changes in conformation; and (5) continue studies on the structure and localization the 14S dynein ATPase. These problems will be addressed by a combination of techniques including: transmission electron microscopic (TEM) localization of monoclonal and polyclonal antibodies; scanning transmission electron microscopy (STEM) of dynein fragments and the microtubule-dynein complex; hydrodynamic and fluorescence anisotropy measurements on isolated dynein; TEM analysis of unstained, unfixed, frozen-hydrated specimens or replicas of rotary shadowed preparations; and solution x-ray scattering studies of the microtubule-dynein complex and intact axonemes. The current work builds upon our previous results on the structure and ATPase pathway of dynein isolated from Tetrahymena cilia. These studies are expected to establish the structure of dynein in detail and to specify the structural basis for changes in conformation necessary to couple ATP hydrolysis to the interaction of the dynein crossbridge with microtubules to produce a force for ciliary movement. The work also provides a basis for analysis of dynein-like ATPases in other microtubule systems such as chromosome movement or the intracellular transport of membrane bound particles.

本研究的总体目标是建立该结构

项目成果

Mechanism of force production for microtubule-dependent movements.

• DOI: 10.1083/jcb.99.1.132s
• 发表时间: 1984-07
• 期刊: The Journal of cell biology
• 作者: Johnson KA;Porter ME;Shimizu T
• 通讯作者: Shimizu T

Structure of the alpha-, beta-, and gamma-heavy chains of 22 S outer arm dynein obtained from Tetrahymena cilia.

从四膜虫纤毛中获得的 22 S 外臂动力蛋白的 α、β 和 γ 重链结构。

• 发表时间: 1989
• 期刊: The Journal of biological chemistry
• 作者: Marchese-Ragona,SP;Facemyer,KC;Johnson,KA
• 通讯作者: Johnson,KA

Structure and mass analysis of 12S and 19S dynein obtained from bull sperm flagella.

从公牛精子鞭毛中获得的 12S 和 19S 动力蛋白的结构和质量分析。

• DOI: 10.1002/cm.970080409
• 发表时间: 1987
• 期刊: Cell motility and the cytoskeleton
• 作者: Marchese-Ragona,SP;Gagnon,C;White,D;Isles,MB;Johnson,KA
• 通讯作者: Johnson,KA