Small Grants for Exploratory Research: Isolation of Myosin-like Organelle Motor

探索性研究的小额资助：肌球蛋白样细胞器运动的分离

• 批准号: 9216634
• 负责人: George Langford
• 金额: $ 4万
• 依托单位: Dartmouth College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9216634&HistoricalAwards=false
• 关键词: Small; Grants; Exploratory; Research; Isolation

The aim of this SGER proposal is to extend a truly novel observation of actin-based motility in axoplasmic extracts. The project calls for the histochemical characterization of the movement system via immunofluorescence and immunogold labelling. S1 decoration will be used to determine actin filament polarity in the context of vectoral particle movement. In addition, the project aims toward the biochemical purification of the motor and its subsequent characterization in terms of its molecular structure, and enzymatic properties in a longer-term aim of establishing its ability to move particles along actin filaments in a totally- characterized system. %%% This project has the potential to alter our perception of axoplasmic particle movement significantly. During the last decade, a growing literature has developed in favor of a microtubule-based motility system for axoplasmic transport. Unidirectional motors that interact with microtubules and operate in anterograde or retrograde particle movement have dominated our thinking about the process. Actin-based motility has received far less attention in this process, though microfilaments have long been known to be present in the axoplasmic extracts that have served as the starting material for the particle movement assays. It is now clear that microfilaments may also serve in particle movement. The clear demonstration that actin-based motility is vectorial, and dependent on the polarity of the actin filaments under physiologically- relevant conditions would be a major contribution to our understanding of cell motility.

这个SGER建议的目的是扩展一个真正新颖的观察肌动蛋白为基础的运动在轴浆提取物。该项目要求通过免疫荧光和免疫金标记对运动系统进行组织化学表征。S1装饰将用于确定矢量粒子运动背景下肌动蛋白丝的极性。此外，该项目旨在对马达进行生化纯化，并根据其分子结构和酶特性对其进行后续表征，以建立其在完全表征的系统中沿肌动蛋白丝移动颗粒的能力。这个项目有可能显著改变我们对轴质粒子运动的感知。在过去的十年中，越来越多的文献支持基于微管的运动系统用于轴浆运输。单向马达与微管相互作用，并在顺行或逆行粒子运动中运行，主导了我们对这一过程的思考。肌动蛋白为基础的运动性在这一过程中受到的关注要少得多，尽管人们早就知道微丝存在于轴浆提取物中，作为颗粒运动测定的起始材料。现在很清楚，微丝也可能在粒子运动中起作用。肌动蛋白为基础的运动是矢量的，并且在生理相关条件下依赖于肌动蛋白丝的极性，这一清楚的证明将对我们对细胞运动的理解做出重大贡献。