Global Analysis of the Rickettsia prowazekii Proteome

立克次体属的成员是流行性和地方性斑疹伤寒和岩石 山和其他斑点发烧。这些疾病在世界范围内构成严重的健康威胁。立克次 流行性斑疹伤寒的病原体普氏菌是一种专性细胞内寄生菌， 只存在于真核宿主细胞的细胞质中。R prowazekii能够利用这种细胞内生态位， 从节肢动物到人类的各种动物，使其能够通过人体虱子传播。这项建议 的蛋白质表达模式的表征。prowazekii。A的重要目标 蛋白质组方法是人类的细菌病原体，表现出独特生物学的生物体， 不幸的是，在目前的人类环境中，潜在的生物恐怖分子。R. prowazekii符合所有三个 此外，由于其小的蛋白质组，这些标准可以用作完整蛋白质组分析的模型。 该建议的具体重点是蛋白质组学技术的发展和应用， 回答关于病原体R的问题。prowazkeii。在具体目标1中，我们的目标是开发和应用高- 全球液相色谱-质谱技术，以获得完整的蛋白质组 被广泛研究的R. prowazeldi马德里E菌株在标准条件下繁殖。我们将应用这个 技术在具体目标2，以确定立克次体蛋白表达的全球模式。立克次体蛋白 将检查在反映该病原体独特生物学的不同条件下表达的病毒。这些 将包括菌株差异、宿主细胞差异、温度变化和导致l_om立克次体的变化。 增长在具体目标3中，我们将通过检查蛋白质组来解决宿主细胞变异的关键延伸 ofR.在节肢动物载体中生长的prowazkeii。这个界面是立克次体存活的关键组成部分。 鉴定立克次体蛋白表达的差异将增加我们对这一动态的理解 这一过程以及确定疫苗和诊断试剂的潜在新靶点。

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 ofa 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 ofR. prowazekii. Important targets for a proteome approach are the bacterial pathogens of humans, organisms that exhibit unique biologies, 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 ofproteomic techniques to address questions about the pathogen R. prowazkeii. 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. prowazeldi 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 l_om rickettsial growth. In Specific Aim 3 we will address a critical extension of host cell variation by examining the proteome ofR. prowazkeii 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.