Biochemical and Functional Analysis of Trypanin

描述（由申请方提供）：非洲锥虫（布氏锥虫）是一种原生动物寄生虫，在人类中引起一种称为非洲昏睡病的致命疾病，在动物中引起相关疾病。这些疾病对健康和经济造成了毁灭性的后果。这些寄生虫通过采采蝇媒介的叮咬传播到哺乳动物宿主的血液中。细胞运动性被怀疑在疾病发病机制和寄生虫发育中起关键作用，但以前不可能直接研究这种关系。我们已经分离出一种锥虫突变体，它不能定向细胞运动，但令人惊讶的是，它没有瘫痪。该突变体缺乏称为“锥虫蛋白”的蛋白质，并且该提议的广泛的、长期的目标是利用锥虫蛋白（-）突变体和重组锥虫蛋白作为工具来推进我们对T.布鲁塞。锥蛋白（-）突变体还将用于研究细胞运动在哺乳动物宿主和寄生虫发育以及舌蝇内疾病发病机制中的作用。前两个具体的目标是确定锥蛋白和锥蛋白相关蛋白（TRP 1）的生化特性，并确定这些蛋白质是否相互作用。胰蛋白酶是T.布鲁氏菌细胞运动性，下一个具体目标是采用组合的生物化学和遗传方法来分离和鉴定该复合物的其它组分。最后，因为锥虫素（-）突变体有一个积极跳动的鞭毛，并没有瘫痪，他们提供了一个独特的机会，专门研究定向细胞运动的作用，在哺乳动物和昆虫宿主。因此，第四个具体目标是确定锥虫蛋白的缺失是否改变哺乳动物宿主中的感染过程和/或阻断采采蝇载体内的寄生虫发育。 T.布氏菌鞭毛附着复合物提供了独特的机会，研究细胞膜上的相互作用的基本方面。因此，除了提供与寄生虫病相关的信息外，这些研究预计与真核细胞生物学具有广泛的相关性。

DESCRIPTION (provided by the applicant): African trypanosomes (Trypanosoma brucei) are protozoan parasites that cause a fatal disease known as African sleeping sickness in humans and related diseases in animals. These diseases have devastating health and economic consequences. These parasites are transmitted to the bloodstream of a mammalian host by the bite of a tsetse fly vector. Cell motility is suspected to play a pivotal role in disease pathogenesis and parasite development, but it has not previously been possible to investigate this relationship directly. We have isolated a trypanosome mutant that is incapable of directional cell motility but, surprisingly, is not paralyzed. This mutant lacks a protein called "trypanin" and the broad, long-term objectives of this proposal are to exploit trypanin (-) mutants and recombinant trypanin proteins as tools to advance our understanding of the mechanism and biology of flagellum based cell motility in T. brucei. Trypanin (-) mutants will also be used to examine the role of cell motility in disease pathogenesis in the mammalian host and parasite development and within the tsetse fly. The first two specific aims are to determine the biochemical properties of trypanin and a trypanin-related protein (TRP 1) and to determine whether these proteins interact with one another. Trypanin is part of a unique flagellum attachment complex that is required for T. brucei cell motility and the next specific aim is to employ a combined biochemical and genetic approach to isolate and identify additional components of this complex. Finally, because trypanin (-) mutants have an actively beating flagellum and are not paralyzed, they provide a unique opportunity to specifically examine the role of directional cell motility in the mammalian and insect hosts. Thus, the fourth specific aim is to determine whether loss of trypanin alters the course of infection in the mammalian host and/or blocks parasite development within the tsetse fly vector. T. brucei flagellum attachment complexes offer unique opportunities to investigate fundamental aspects of cytoskeleton-membrane interactions. Thus, in addition to providing information relevant to parasitic disease, these studies are expected to have broad relevance to eukaryotic cell biology.