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

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 轴突运输是由属于动蛋白和细胞质动力蛋白超家族的马达蛋白驱动的膜结合颗粒沿微管运输的过程。最近，我们发现了一种与轴浆细胞器协同纯化并定位于细胞器/微管界面的激动素-3(KIF1A)。针对Kinesin-3的抗体修饰挤出的轴浆中的100 nm细胞器，并抑制细胞器向微管正端的运输。这些发现表明，Kinesin-3可能负责将100 nm的细胞器移动到微管的正端。在这里，我们建议表征纯化的Kinesin-3的运动特性，并鉴定Kinesin-3货物的分子解剖学。了解Kinesin-3的运动速度和方向及其产物的分子组成将有助于确定Kinesin-3在轴突中的作用，并确定其在运动介导的运输中的作用。

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.