GENETIC AND IMMUNOLOGIC ANALYSIS OF MICROTUBULE PROTEINS

驱动蛋白及其亲属是微管马达， 通过将ATP中的化学能转化为 力和运动。虽然对这些组织了解得很多， 电动机及其潜在功能，我们的知识中仍然存在许多漏洞。 特别是，我们不明白机械化学转导 我们也不完全了解驱动蛋白马达是如何 在细胞中调节和发挥作用。 本提案中所述工作的目标是阐明这些 实现几个相互关联的目标。第一，扩大我们的 了解机械化学力传递，我们将获得高- 质量晶体的驱动蛋白马达域，并解决其原子 结构其次，在驱动蛋白运动的补充分析中，我们 将识别与微管结合相关的区域， 机械化学中重要的序列。第三，扩大我们的 了解驱动蛋白的调节和功能，我们将重点关注 在驱动蛋白上相互作用的蛋白质的遗传和生物化学分析 尾部，特别是在驱动蛋白轻链和驱动蛋白上，其可以 是参与驱动蛋白功能的调节或附着蛋白。 第四，扩大我们对轴突运输机制的认识， 驱动蛋白马达所扮演的角色，我们将从基因上分析 KLP 64 D和KLP 68 D的功能，这是两个驱动蛋白的亲戚， 很可能在顺行轴突运输中起作用，沿着 驱动蛋白总之，我们的研究应该大大促进我们的 了解驱动蛋白的功能和功能机制 及其亲属。

Kinesin and its relatives are microtubule motors that generate intracellular movements by converting the chemical energy in ATP into force and movement. While much is known about the organization of these motors and their potential functions, many holes in our knowledge remain. In particular, we do not understand how the mechanochemical transduction of ATP occurs, nor do we fully understand how kinesin motors are regulated and function in the cell. The goal of the work described in this proposal is to elucidate these issues by achieving several interrelated goals. First, to expand our understanding of mechanochemical force transduction, we will obtain high- quality crystals of the kinesin motor domain and solve its atomic structure. Second, in complementary analyses of the kinesin motor, we will identify regions involved in binding to the microtubule, and sequences important in mechanochemistry. Third, to broaden our understanding of kinesin regulation and function, we will focus on genetic and biochemical analysis of proteins that interact at the kinesin tail, in particular on the kinesin light chains and kinectin, which may be regulatory or attachment proteins involved in kinesin function. Fourth, to extend our knowledge of the mechanisms of axonal transport and the roles played by kinesin motors, we will genetically analyze the functions of KLP64D and KLP68D, which are two kinesin relatives that are very likely to function in anterograde axonal transport along with kinesin. Taken together, our studies should significantly advance our understanding of the functions, and mechanisms of function, of kinesin and its relatives.