Global Analysis of the Rickettsia prowazekii Proteome

描述（由申请人提供）：立克次体属的成员是流行和地方性斑疹伤寒、落基山热和其他斑疹热的病原。这些疾病在世界范围内对健康构成重大威胁。普氏立克次体是流行性斑疹伤寒的病原，是一种专性细胞内寄生细菌，只能在真核宿主细胞的细胞质内生长。prawazekii能够利用这种细胞内的生态位在动物身上传播，包括节肢动物和人类，从而使其成为人体虱子的载体。本研究主要是对propraazekii的蛋白表达模式进行研究。蛋白质组学方法的重要目标是人类的细菌病原体，具有独特生物学特性的生物体，不幸的是，在当前的人类环境中，潜在的生物恐怖分子。r.p awazekii符合所有这三个标准，此外，由于其小蛋白质组，可以作为完整蛋白质组分析的模型。本建议的重点是蛋白质组学技术的发展和应用，以解决有关病原体的问题。在具体目标1中，我们的目标是开发和应用高通量，全球液相色谱-质谱技术，以获得在标准条件下繁殖的广泛研究的R. prowazekii Madrid E菌株的完整蛋白质组。我们将在Specific Aim 2中应用这项技术来确定立克次体蛋白表达的全球模式。立克次体蛋白在不同条件下表达，反映这种病原体的独特生物学将被检查。这些将包括菌株差异、宿主细胞差异、温度变化以及由立克次体生长引起的变化。在Specific Aim 3中，我们将通过检测在节肢动物载体中生长的proproazekii的蛋白质组来解决宿主细胞变异的关键扩展。这个界面是立克次体存活的关键组成部分。确定立克次体蛋白表达的差异将增加我们对这一动态过程的理解，并确定疫苗和诊断试剂的潜在新靶点。

DESCRIPTION (provided by applicant): Members of the genus Rickettsia are the etiologic agents of epidemic and endemic typhus and rocky mountain and other spotted fevers. These diseases pose significant health threats worldwide. Rickettsia prowazekii, the etiologic agent of epidemic typhus, is an obligate intracellular parasitic bacterium that can grow only within the cytoplasm of a eukaryotic host cell. R prowazekii is able to exploit this intracellular niche in animals as diverse as arthropods and humans, allowing it to be vectored by the human body louse. This proposal is focused on the characterization of the protein expression patterns of R. prowazekii. Important targets for a proteome approach are the bacterial pathogens of humans, organisms that exhibit unique biology's, and unfortunately, in the current human environment, potential bioterrorist agents. R. prowazekii fits all three of these criteria and in addition, due to its small proteome, can serve as a model for complete proteome analysis. The specific focus of this proposal is on the development and application of proteomic techniques to address questions about the pathogen R. prowazekii. In Specific Aim 1 our goal is to develop and apply high-throughput, global liquid chromatography-mass spectrometry technology to obtain the complete proteome of the extensively studied R. prowazekii Madrid E strain propagated under standard conditions. We will apply this technology in Specific Aim 2 to identify global patterns of rickettsial protein expression. Rickettsial proteins expressed under different conditions that reflect the unique biology of this pathogen will be examined. These will include strain differences, host cell differences, temperature variation, and changes resulting from rickettsia growth. In Specific Aim 3 we will address a critical extension of host cell variation by examining the proteome of R. prowazekii growing in an arthropod vector. This interface is a critical component of rickettsial survival. Identification of differences in rickettsial protein expression will increase our understanding of this dynamic process as well as identify potentially new targets for vaccines and diagnostic reagents.