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; 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）：立克属的成员是流行病和地方性斑疹伤寒和落基山的病因学药物以及其他斑点发烧。这些疾病在全球构成了重大健康威胁。 Rickettsia prowazekii是流行病的病因学剂，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中生长。 R Prowazekii能够利用像节肢动物和人类一样多样化的动物中这种细胞内生态位，从而使其被人体虱子载入。该建议的重点是prowazekii的蛋白质表达模式的表征。蛋白质组方法的重要靶标是人类的细菌病原体，具有独特生物学的生物，不幸的是，在当前的人类环境中，潜在的生物恐怖剂。 R. prowazekii适合所有这三个标准，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。该提案的具体重点是蛋白质组学技术的开发和应用，以解决有关病原体R. prowazekii的问题。在特定目的1中，我们的目标是开发和应用高通量，全球液态色谱 - 质谱技术，以获取在标准条件下传播的广泛研究的广泛研究的全蛋白质组。我们将在特定的目标2中应用这项技术，以确定立克蛋白表达的全球模式。将检查反映该病原体独特生物学的不同条件下表达的立克蛋白。这些将包括应变差异，宿主细胞差异，温度差异以及身心重量的变化。在特定的目标3中，我们将通过检查节肢动物载体中生长的R. prowazekii的蛋白质组来解决宿主细胞变异的关键扩展。该界面是立克西亚生存的关键组成部分。鉴定立克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标。