A NOVEL MICROTUBULE-ASSOCIATED PROTEIN IN TRYPANOSOMES

描述（改编自应用摘要）：非洲锥虫是 原生动物寄生虫，导致非洲锥虫病，一种致命的疾病， 破坏性的健康和经济后果。的长期目标 这项研究的目的是为了加深我们对两个重要组成部分的理解， 这些病原体的细胞结构：微管细胞骨架， 鞭毛袋是一种特殊的细胞器， 宿主与寄生虫之间的交流这两种结构都需要 寄生虫在其哺乳动物宿主和昆虫载体中存活， 被认为是治疗干预的主要目标， 锥虫我们已经证明，一种名为TLTF的蛋白质定位于 锥虫前鞭毛袋膜的胞质面 蛋白质提取物和一种相关的人类蛋白质与微管结合， 哺乳动物细胞建议研究的前两个目标是利用 结合体内和体外微管结合试验，以确定是否 这两种蛋白是真正的微管结合蛋白， 确定微管是否有助于靶向TLTF， 锥虫这项研究的另一个目的是利用生物化学和分子生物学技术， 生物学方法来确定两个抗- TLTF交叉反应蛋白，存在于昆虫形式的锥虫中。 这些蛋白质之一的表达表现出戏剧性的发育 调节，表明该蛋白具有发育阶段特异性 功能协调发展的TLTF靶向结构域中的点突变分析表明， 鞭毛袋中特定的蛋白质间相互作用 这对TLTF的正确定位很重要。因此，本研究的另一个目的是 使用免疫共沉淀和亲和捕获树脂来确定是否 TLTF与其他锥虫蛋白相互作用，即是否有些TLTF “结合配偶体”可以是鞭毛袋的组分。除了 对我们理解医学上重要的细胞生物学的贡献 病原体，TLTF的密切相关的性质和以前的 这些研究的结果可能是， 将在其他系统中具有广泛的生物相关性。

DESCRIPTION (adapted from application abstract): African trypanosomes are protozoan parasites that cause African trypanosomiasis, a deadly disease with devastating health and economic consequences. The long-term goal of the proposed research is to advance our understanding of two vital components of the cellular architecture of these pathogens: the microtubule cytoskeleton, and the flagellar pocket, a specialized organelle that plays a critical role in host-parasite communication. Both of these structures are required for parasite survival in their mammalian host and insect vector, and are considered to be prime targets for therapeutic intervention in trypanosomiasis. We have shown that a protein called TLTF is localized to the cytoplasmic face of the anterior flagellar pocket membrane in trypanosome protein extracts and a related human protein associates with microtubules in mammalian cells. The first two aims of the proposed research are to utilize combined in vivo and in vitro microtubule binding assays to determine whether these two proteins are bona fide microtubule-binding proteins, and to determine whether or not microtubules contribute to targeting of TLTF in trypanosomes. Another aim of this research is to use biochemical and molecular biological approaches to determine whether the relationship between two anti- TLTF cross-reactive proteins that are present in insect-form trypanosomes. Expression of one of these proteins exhibits dramatic developmental regulation, suggesting that this protein has developmental stage-specific functions. Analysis of point mutations in the TLTF targeting domain suggests that specific protein-protein interactions at the flagellar pocket are important for proper targeting of TLTF. Hence, another aim of this research is to use co-immunoprecipitation and affinity capture resins to determine whether TLTF interacts with other trypanosomal proteins, i.e, whether some TLTF `binding partners` may be components of the flagellar pocket. In addition to contribution to our understanding of the cell biology of a medically important pathogen, the closely related properties of TLTF and a previously uncharacterized human protein make it likely that findings from these studies will have broad biological relevance in other systems.