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: 6672487
• 负责人: DAVID WOOD
• 金额: $ 17.96万
• 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/6672487
• 关键词: Rickettsia prowazekii; Rickettsiales disease; Siphonaptera; bacterial genetics; bacterial proteins; disease vectors; genetic strain; high throughput technology; host organism interaction; liquid chromatography mass spectrometry; microorganism population study; protein biosynthesis; proteomics; technology /technique development; temperature; tissue /cell culture

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（由申请人提供）：立克次体属的成员是流行性和地方性斑疹伤寒、落基山热和其他斑点热的病原体。这些疾病在世界范围内构成严重的健康威胁。普氏立克次体是一种专性胞内寄生菌，只能在真核宿主细胞的细胞质内生长，是引起流行性斑疹伤寒的病原体。R prowazekii能够在节肢动物和人类等多种动物中利用这种细胞内生态位，使其能够通过人体虱子传播。本研究的重点是对R. prowazekii。蛋白质组方法的重要目标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，在当前的人类环境中，潜在的生物恐怖分子。R. prowazekii符合所有这三个标准，此外，由于其小的蛋白质组，可以作为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，以解决有关病原体R。prowazekii。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，以获得广泛研究的R。prowazekii马德里E菌株在标准条件下繁殖。我们将在特定目标2中应用该技术来识别立克次体蛋白表达的全球模式。将检查在不同条件下表达的反映该病原体独特生物学的立克次体蛋白。这些将包括菌株差异、宿主细胞差异、温度变化和立克次体生长引起的变化。在具体目标3中，我们将通过研究R. prowazekii在节肢动物载体中生长。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过程的理解，并为疫苗和诊断试剂确定潜在的新靶点。