CYTOPLASMIC GELATION AND CELL MOVEMENT IN DICTYOSTELIUM

网网柄菌中的细胞质凝胶化和细胞运动

• 批准号: 3288156
• 负责人: Marcus Fechheimer
• 金额: $ 9.41万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-10 至 1988-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3288156
• 关键词: antibody formation; cell motility; chemical binding; cytoplasm; cytoskeleton; electron microscopy; enzyme linked immunosorbent assay; fluorescence microscopy; gel; gel electrophoresis; image processing; immunofluorescence technique; ion exchange chromatography; molecular biology; molecular weight; monoclonal antibody; phagocytosis; stoichiometry

Cell movements are essential to the existence of essentially all eucaryotic cells. These movements must involve the interaction of force generating and structural constituents of cells. The force for many cell movements is thought to be generated by actin and myosin, two proteins known to participate in muscle contraction. Structural components of cells that may participate in cell movements have recently been discovered in a number of cell types. These proteins form structured gels when mixed with actin in vitro. Actin-binding proteins from Dictyostelium discoideum having subunit molecular weights of 30,000 and 95,000 daltons will be examined in this proposal. First, the hypothesis that these components are required for the movement of living cells will be tested. Monoclonal antibodies that bind to and inhibit the activity of these structural proteins will be prepared and incorporated into living cells. Measurements of the movement of those cells will test the requirement for the specific proteins. Second, the structure and actin binding activities of the 30,000 and 95,000 dalton proteins will be examined in detail using viscometry, sedimentation, and ultrastructural techniques. Third, the presence of the 30,000 dalton protein in cellular extensions, pseudopodia and filopodia, will be assessed using both biochemical and immunochemical approaches. The results will both test and extend our knowledge of the molecular mechanisms of cell movements. Since the cytoskeletal and contractile proteins of D. discoideum are homologous to those in both invertebrate and vertebrate cell types, study of their interactions and intracellular functions may have general significance. The panoramic importance of cell movements is verified by their prominence in developmental biology, cellular immunology, and oncology. Knowledge of the molecular mechanisms of cell movements will direct future studies of these phenomena in normal and pathological states, and may eventually contribute to development of therapeutic and diagnostic procedures.

细胞运动对于基本上所有真核生物的存在都是必不可少的。 细胞 这些运动必须涉及力的相互作用， 和细胞的结构成分。 许多细胞运动的动力是 被认为是由肌动蛋白和肌球蛋白，两种蛋白质已知， 参与肌肉收缩。 细胞的结构成分， 参与细胞运动最近被发现在一些 细胞类型。 当这些蛋白质与肌动蛋白混合时， 体外 具有亚基的盘基网柄藻肌动蛋白结合蛋白 在此将检查30，000和95，000道尔顿的分子量。 提议 首先，假设这些组件是必需的， 将测试活细胞的运动。 结合的单克隆抗体 并抑制这些结构蛋白的活性 并整合到活细胞中。 测量那些 细胞将测试对特定蛋白质的需求。 二是 30，000和95，000道尔顿的结构和肌动蛋白结合活性 蛋白质将使用粘度法、沉降法和 超微结构技术 第三，30，000道尔顿的存在 将评估细胞延伸物伪足和丝状伪足中的蛋白质 使用生物化学和免疫化学方法。 结果将 这两个测试和扩展我们的知识的分子机制的细胞 动作 由于D. 盘状突与无脊椎动物和脊椎动物细胞中盘状突同源 类型，它们的相互作用和细胞内功能的研究可能有 一般意义。 细胞运动的重要性在于 通过它们在发育生物学，细胞免疫学， 和肿瘤学 细胞运动的分子机制的知识将 指导未来在正常和病理状态下对这些现象的研究， 并可能最终有助于开发治疗和诊断 程序.