STRUCTURAL STUDIES ON MOLECULAR TRACKS AND MOTORS

分子轨道和马达的结构研究

• 批准号: 6497408
• 负责人: RONALD A MILLIGAN
• 金额: $ 30.63万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6497408
• 关键词: Acanthamoeba; Dictyostelium; actins; calmodulin; cell motility; chickens; conformation; cryoelectron microscopy; dynein ATPase; gold; image processing; imaging /visualization /scanning; laboratory rabbit; molecular dynamics; myosins; phosphorylation; protein structure function; smooth muscle; structural biology

DESCRIPTION: The interactions of molecular tracks and motors result in movements in cells. Fundamental to our understanding of the way in which motility is accomplished in cells is a detailed description not only of the atomic structures of the complexes involved, but also the way in which the activity of these complexes is regulated in response to the needs of the cell. The long term goals of the work described in this application are to understand in structural terms, the way in which myosin motors are regulated. Three types of myosin-based regulation will be investigated: (I) Regulation mediated by light chain phosphorylation will be investigated by examining a number of smooth muscle myosin constructs. (ii) Regulation by light chain dissociation will be examined in brush border myosin l and myosin V. (iii) Regulation by heavy chain phosphorylation will be investigated in Acanthamoeba and Dictyostelium myosin ls. The methods to be used include cryoelectron microscopy, helical image analysis and modeling using the x-ray structures of the myosin head and regulatory domain. The experiments have been designed to visualize actomyosin conformations which are the basis of the regulatory mechanism. Atomic models of the regulated and activated states will be built from the EM and X-ray data. The results should provide detailed insights into the structural basis for regulation in myosin involved in uterine contractions, blood pressure maintenance and intestinal peristalsis (smooth muscle myosin), and in intracellular trafficking and endocytosis (myosin ls and Vs). Some exploratory experiments on dynein are planned.

描述：分子轨道和马达的相互作用导致 细胞内的运动。 对我们理解方法的基础 运动是在细胞中完成的，这不仅是对细胞的详细描述 所涉及的配合物的原子结构，以及其中的方式 这些复合物的活性根据需要进行调节 细胞。 本申请中描述的工作的长期目标是 从结构角度理解肌球蛋白马达的运作方式 受监管。 将研究三种类型的基于肌球蛋白的调节：（I）调节 通过检查轻链磷酸化介导的 平滑肌肌球蛋白结构的数量。 (ii) 轻链调节 将检查刷状缘肌球蛋白 L 和肌球蛋白 V 的解离。 (iii) 重链磷酸化的调节将在以下方面进行研究 棘阿米巴和盘基网柄菌肌球蛋白 ls。 使用的方法包括冷冻电子显微镜、螺旋图像 使用肌球蛋白头部的 X 射线结构进行分析和建模 监管领域。 实验旨在可视化 肌动球蛋白构象是调节机制的基础。 调节状态和激活状态的原子模型将根据 EM 和 X 射线数据。 结果应提供详细的见解 参与子宫收缩的肌球蛋白调节的结构基础， 血压维持和肠道蠕动（平滑肌 肌球蛋白），以及细胞内运输和内吞作用（肌球蛋白 ls 和 VS）。 计划对动力蛋白进行一些探索性实验。