GENETIC AND IMMUNOLOGIC ANALYSIS OF MICROTUBULE PROTEINS

真核细胞使用基于微管的转运来完成 胞内细胞器运动，细胞分裂，可能还有细胞 形态发生。这些事件需要微管马达的活动。 蛋白质，如激动素。在这个提案中，我们描述了实验 旨在了解动蛋白如何将化学能转化为 机械力，动蛋白如何附着在它运动的元素上， 激动素的哪些结构特征是体内功能所必需的， 当Kinesin符合所有基于MT的运动方案时，一些 这可能是由类似动蛋白的马达产生的。要实现这些目标 目标，我们将：1)继续我们对动蛋白元素的研究 力产生所需的重链电机领域及开发方法 用于大规模生产马达领域的Kinesin 高分辨率结构研究；2)开始分析蛋白质 运动蛋白重链尾巴相互作用以连接到 蜂窝货物。3)以确定的方式改变肌动蛋白重链和 将这些改变的分子重新引入生物体中进行研究 以及4)分析我们最近发现的五个新基因 似乎编码了激动素样蛋白。总之，我们的调查将 揭示一个特定的微管马达是如何工作的，并将开始 解开真核生物用来 完成基于微管的运动。

Eukaryotic cells use microtubule-based transport to accomplish intracellular organelle movement, cell division, and possibly cellular morphogenesis. These events require the activities of microtubule-motor proteins such as kinesin. In this proposal we describe experiments designed to understand how kinesin converts chemical energy into mechanical force, how kinesin attaches to the elements that it moves, what structural features of kinesin are essential for in vivo function, and where kinesin fits in the scheme of all MT-based movements, some of which might be generated by kinesin-like motors. To accomplish these goals, we will: 1) continue our studies of the elements of the kinesin heavy chain motor domain needed for force generation and develop methods for large-scale production of the kinesin motor domain for use in high-resolution structural studies; 2) begin analyses of proteins with which the kinesin heavy chain tail interacts in order to attach to cellular cargoes. 3) alter kinesin heavy chain in defined ways and introduce these altered molecules back into the organism for study in vivo; and 4) analyze five new genes that we have recently discovered that appear to encode kinesin-like proteins. In toto, our investigations will reveal how one particular microtubule motor functions and will begin to unravel the range of functions and strategies used by eukaryotes to accomplish microtubule-based movements.