Global Analysis of the Rickettsia prowazekii Proteome

普瓦泽基立克次体蛋白质组的整体分析

• 批准号: 6754378
• 负责人: DAVID WOOD
• 金额: $ 36.22万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6754378
• 关键词: Global; Analysis; Rickettsia; prowazekii; Proteome

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.

描述(申请人提供)：立克次体属成员是流行性和地方性斑疹伤寒、落基山斑疹伤寒和其他斑点热病的病原体。这些疾病在世界范围内构成了重大的健康威胁。普氏立克次体是流行性斑疹伤寒的病原体，是一种专性的细胞内寄生细菌，只能在真核宿主细胞的细胞质内生长。R prowazekii能够利用节肢动物和人类等各种动物的这种细胞内生态位，使其能够通过人体虱子进行传播。本研究的重点是对普罗瓦泽克乳杆菌的蛋白质表达模式进行研究。蛋白质组方法的重要目标是人类的细菌病原体，即表现出独特生物学特征的生物体，不幸的是，在目前的人类环境中，潜在的生物恐怖分子。Prowazekii菌株符合上述三个标准，此外，由于其蛋白质组较小，可以作为完整蛋白质组分析的模型。这项提案的具体重点是开发和应用蛋白质组学技术来解决关于病原体R.prowazekii的问题。在具体目标1中，我们的目标是开发和应用高通量、全局液-质联用技术，以获得在标准条件下繁殖的被广泛研究的普氏弧菌马德里E株的完整蛋白质组。我们将把这项技术应用于特定的目标2，以确定立克次体蛋白表达的全球模式。将检测在不同条件下表达的反映该病原体独特生物学特性的立克次体蛋白。这将包括菌株差异、宿主细胞差异、温度变化和立克次体生长引起的变化。在特定的目标3中，我们将通过研究生长在节肢动物载体中的普氏根结线虫的蛋白质组来解决宿主细胞变异的关键扩展。这种界面是立克次体生存的关键组成部分。识别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，并确定潜在的疫苗和诊断试剂的新靶点。