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

这项SGER提案的目的是扩展一个真正新颖的 观察轴浆提取物中基于肌动蛋白的运动性。的 该项目要求组织化学表征的 运动系统通过免疫荧光和免疫金标记。S1 装饰将被用来确定肌动蛋白丝极性在 矢量粒子运动的背景。该项目还 目的是对发动机进行生化净化， 随后根据其分子结构进行表征， 和酶的性质，在建立其长期目标， 使粒子沿着肌动蛋白丝运动的能力， 特色系统 %%% 这个项目有可能改变我们对 轴浆颗粒运动明显。在过去的十年里， 越来越多的文献支持基于微管的 轴浆运输动力系统。单向电动机 与微管相互作用， 逆行粒子运动主导了我们对 过程肌动蛋白为基础的运动性受到的关注要少得多， 这个过程，虽然微丝早已被认为是 存在于轴浆提取物中， 用于颗粒运动测定的材料。现在已经清楚 微丝也可用于颗粒运动。的明确 证明肌动蛋白为基础的运动是矢量的，依赖于 在生理条件下肌动蛋白丝的极性 相关条件将是我们的一个重大贡献， 对细胞运动的理解。

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.