CYTOCHALASINS, ACTIN FILAMENTS, AND CELL MOTILITY

细胞松弛素、肌动蛋白丝和细胞运动

• 批准号: 3271059
• 负责人: Shin Lin
• 金额: $ 20.3万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-05-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3271059
• 关键词: affinity chromatography; autoradiography; cell biology; cell growth regulation; cell motility; chemical binding; cytochalasins; cytoskeleton; electron spin resonance spectroscopy; erythrocyte membrane; erythrocytes; fluorescence microscopy; human tissue; immunochemistry; membrane reconstitution /synthesis; platelets; polymerization; radiotracer; tissue /cell culture

The long-term goal of this project is to determine the molecular basis of the inhibitory effects of cytochalasins (a group of fungal metabolites) on a wide spectrum of cytoskeletal and contractile functions in animal cells and tissues. The results to be obtained from this study will enable us to use these drugs effectively as diagnostic tools in future biomedical research. Moreover, this study is also aimed at yielding information on the molecular mechanisms by which a cell controls its motile functions. It has previously been shown that cytochalasins have the ability to interact with actin, a major protein component of the cytoskeleton of nonmuscle cells. In the proposed research, we will continue our use of a combination of biochemical, biophysical, and cell biological techniques to probe at the action of cytochalasins and related protein factors on actin structure and function in the following ways. (A) The high-affinity and low-affinity binding of different cytochalasins to monomeric and polymeric actin will be measured with standard pharmacological techniques. The binding data will then be correlated with studies on the effects of the drugs on actin filament assembly, disassembly, and network formation in vitro, under defined conditions and using purified protein components. (B) The efects of cytochalasins on actin-containing structures in cellylar and subcellular systems will be determined. This information will be related to the concentration dependency of the inhibition of a number of motile functions by cytochalasins, and to the data obtained in the in vitro experiments described above. (C) The structure of actin-containing cytochalasin binding complexes isolated from motile cells (e.g. human platelets) will be studied. Experiments will also be conducted to investigate the mechanism by which these complexes associate with cellular membranes. (D) Comparative studies on a class of proteins with cytochalasin-like activity against actin filament assembly and interactions will be continued. Special emphasis will be directed at the possibility that these proteins may function as regulators of actin filament assembly and membrane attachment in the cell.

这个项目的长期目标是确定分子基础 细胞松弛素(一组真菌代谢产物)的抑制作用 动物细胞中广泛的细胞骨架和收缩功能 还有纸巾。这项研究的结果将使我们能够 在未来的生物医学中有效地使用这些药物作为诊断工具 研究。此外，这项研究还旨在获得关于 细胞控制其运动功能的分子机制。它 先前已经证明，细胞松弛素具有相互作用的能力 肌动蛋白，非肌肉细胞骨架的主要蛋白质成分 细胞。在拟议的研究中，我们将继续使用组合 生化、生物物理和细胞生物学技术在 细胞松弛素及相关蛋白因子对肌动蛋白结构和功能的影响 通过以下方式发挥作用。(一)高亲和力与低亲和力 不同的细胞松弛素与单体和聚合肌动蛋白的结合将是 用标准的药理学技术测量。绑定数据将 然后与药物对肌动蛋白影响的研究相关联 体外纤维组装、分解和网络形成，在 确定的条件和使用纯化的蛋白质组分。(B)影响 细胞松弛素对细胞和亚细胞内含肌动蛋白结构的影响 系统将被确定。此信息将与 多项运动功能抑制的浓度依赖性 通过细胞松弛素，以及在体外实验中获得的数据 如上所述。(C)含肌动蛋白的细胞松弛素的结构 从运动细胞(例如人血小板)中分离出来的结合复合体将是 学习。还将进行实验来研究这种机制。 这些复合体通过它与细胞膜相联系。(D) 一类具有细胞松弛素样活性的蛋白质的比较研究 针对肌动蛋白细丝的组装和相互作用将继续进行。 特别强调的是这些蛋白质可能 可作为肌动蛋白细丝组装和膜的调节器 附着在细胞中。