CHARACTERIZATION OF KINESIN-3 & CARGO IN TRANSIT WITHIN THE SQUID GIANT AXON

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 轴突运输是一个过程，其中膜结合的颗粒是在运输沿着微管动力属于驱动蛋白和细胞质动力蛋白超家族的马达蛋白。最近，我们发现了一个驱动蛋白-3（KIF 1A），它与轴浆细胞器共纯化，并定位于细胞器/微管界面。针对驱动蛋白-3产生的抗体在挤出的轴浆中装饰100 nm的细胞器，并抑制细胞器向微管的正端运输。这些发现表明，驱动蛋白-3可能是负责移动100纳米的细胞器向正端的微管。在这里，我们提出了纯化的驱动蛋白-3的电机特性的特点，并确定驱动蛋白-3货物的分子解剖。了解驱动蛋白3运动的速率和方向及其货物的分子组成将有助于确定驱动蛋白3在轴突中的作用，并确定其对运动介导的运输的贡献。

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. Axonal transport is a process in which membrane bound particles are in transit along microtubules powered by motor proteins belonging to the kinesin and cytoplasmic dynein superfamilies. Recently we identified a Kinesin-3 (KIF1A) that co-purifies with axoplasmic organelles and localizes to organelle/microtubule interfaces. Antibodies raised against Kinesin-3 decorate 100 nm organelles in extruded axoplasm and inhibit organelle transport towards the plus ends of microtubules. These finding suggest that Kinesin-3 may be responsible for moving 100 nm organelles towards the plus ends of microtubules. Here, we propose to characterize the motor properties of purified Kinesin-3 and to identify the molecular anatomy of the Kinesin-3 cargo. Understanding the rate and direction of Kinesin-3 movement and the molecular composition of its cargo will help establish the role of Kinesin-3 in the axon and determine it's contribution to motor mediated transport.