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: 7157623
• 负责人: DAVID WOOD
• 金额: $ 34.61万
• 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 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7157623
• 关键词: Address; Animals; Antibiotics; Arthropod Vectors; Arthropods; Bacteria; Biological Models; Biology; Cells; Complement; Condition; Cytoplasm; Development; Diagnostic Reagent; Disease; Egg Yolk; Environment; Epidemic; Exhibits; Fever; Fractionation; Genes; Genome; Goals; Gold; Growth; Health; Human; Human body; Investigation; Mass Spectrum Analysis; Metabolic; Methods; Modeling; Organism; Pattern; Pediculus humanus humanus; Physiology; Procedures; Process; Proteins; Proteome; Protocols documentation; Rickettsia; Rickettsia prowazekii; Spottings; Standards of Weights and Measures; Stress; Techniques; Technology; Temperature; Typhus; Vaccines; Variant; Work; Yolk Sac; base; design; egg; intracellular parasitism; liquid chromatography mass spectrometry; member; novel; pathogen; protein expression; tissue/cell culture

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.

立克次体属的成员是流行性和地方性斑疹伤寒和石斑疹的病原体。 高山和其他斑点发烧。这些疾病在世界范围内构成了重大的健康威胁。立克次体 流行性斑疹伤寒的病原体是一种专性的细胞内寄生细菌，可以生长。 仅存在于真核宿主细胞的细胞质内。R prowazekii能够利用这种细胞内生态位 各种动物，如节肢动物和人类，使其能够通过人体虱子传播。这项建议 重点是对OFR的蛋白质表达模式的表征。普罗瓦泽基。重要的目标 蛋白质组方法是人类的细菌病原体，表现出独特生物学特性的生物体，以及 不幸的是，在目前的人类环境中，潜在的生物恐怖分子。PROWAZEKII符合所有三个 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蛋白质组技术的开发和应用 回答有关病原体R.prowazkeii的问题。在具体目标1中，我们的目标是开发和应用高 吞吐量，全球液-质联用技术获得完整的蛋白质组 被广泛研究的R.prowazeldi马德里E株在标准条件下繁殖。我们将应用这一点 特定目标2中的技术，以确定立克次体蛋白表达的全球模式。立克次体蛋白 将检查在不同条件下表达的反映该病原体独特生物学特性的基因。这些 将包括菌株差异、宿主细胞差异、温度变化和由L立克次体引起的变化 成长。在特定的目标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扩展 OFR。Prowazkeii生长在节肢动物媒介中。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种动态的理解 以及确定疫苗和诊断试剂的潜在新目标。