PROTEIN COMPOSITION OF ACTIN TAILS ASSEMBLED IN YEAST EXTRACTS

酵母提取物中组装的肌动蛋白尾部的蛋白质组成

• 批准号: 8365818
• 负责人: DAVID G DRUBIN
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365818
• 关键词: Actins; Biology; Cells; Clathrin; Complex; Cytosol; Defect; Disease; Endocytosis; Environment; Funding; Fungal Genome; Grant; Laboratories; Life; Mass Spectrum Analysis; Mediating; Membrane; Microfilaments; Microspheres; Molecular; National Center for Research Resources; Polystyrenes; Principal Investigator; Process; Proteins; Reaction; Research; Research Infrastructure; Resources; Role; Saccharomyces cerevisiae; Saccharomycetales; Set protein; Source; Structure; System; Tail; United States National Institutes of Health; Vesicle; Yeasts; base; coated pit; cost; extracellular; in vivo; macromolecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Clathrin-mediated endocytosis is a fundamental process in living cells, allowing them to capture macromolecules efficiently from the extracellular environment and package them into vesicles in the cytosol. Numerous diseases can be traced to a defect in the succession of molecular reactions leading to endocytosis. In the budding yeast Saccharomyces cerevisiae, endocytosis is localized to cortical actin patches, where a dense network of actin filaments is polymerized around clathrin-coated pits. This network is regulated by a complex set of proteins which cooperate in order to generate membrane invagination. We have recently developed in our laboratory a system in yeast extracts for the assembly of actin based structures around polystyrene microbeads. We believe that the protein composition of these structures could be similar to the protein composition found in vivo in cortical actin patches. We isolated the beads with their associated actin structures, and we would like to determine the protein composition by using mass spectroscopy. These results should allow us to further analyze the role of the different components during endocytosis.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 网格蛋白介导的内吞作用是活细胞中的一个基本过程，使它们能够从细胞外环境中有效地捕获大分子并将其包装到细胞质中的囊泡中。许多疾病都可以追溯到导致内吞作用的一系列分子反应的缺陷。在出芽的酿酒酵母中，内吞作用局限于皮质肌动蛋白斑块，其中肌动蛋白丝的致密网络在网格蛋白包被的小坑周围聚合。该网络由一组复杂的蛋白质调节，这些蛋白质相互协作以产生膜内陷。 我们最近在实验室开发了一种酵母提取物系统，用于在聚苯乙烯微珠周围组装基于肌动蛋白的结构。我们相信这些结构的蛋白质组成可能与体内皮质肌动蛋白斑块中发现的蛋白质组成相似。我们分离了珠子及其相关的肌动蛋白结构，并且我们希望通过使用质谱法来确定蛋白质组成。这些结果应该使我们能够进一步分析不同成分在内吞作用过程中的作用。