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.

立克属的成员是流行病和地方性斑疹伤寒和岩石的病因学剂 山和其他斑点发烧。这些疾病在全球构成了重大健康威胁。人力车 Prowazekii是流行病的病因学剂，是一种可强的细胞内寄生细菌 仅在真核宿主细胞的细胞质内。 r prowazekii能够利用这种细胞内的利基市场 像节肢动物和人类一样多样化的动物，使它被人体虱子载入。这个建议 专注于蛋白质表达模式的表征。 prowazekii。一个重要目标 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，以及 不幸的是，在当前的人类环境中，潜在的生物恐怖剂。 R. Prowazekii适合所有三个 这些标准及其由于其小蛋白质组而言，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技术的开发和应用 解决有关病原体R. Prowazkeii的问题。在特定目的1中，我们的目标是开发和应用高度 - 吞吐量，全球液相色谱 - 质谱技术，以获得完整的蛋白质组 广泛研究了在标准条件下繁殖的prowazeldir。prowazeldie菌株。我们将应用此 特定目标2的技术以识别立克蛋白表达的全球模式。立克蛋白 将在不同条件下反映这种病原体的独特生物学。这些 将包括应变差异，宿主细胞差异，温度变化以及导致L_OM立克的变化 生长。在特定目标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。在节肢动物载体中生长的prowazkeii。该界面是立克西亚生存的关键组成部分。 立克蛋白表达差异的识别将增加我们对这种动态的理解 过程以及确定疫苗和诊断试剂的潜在新靶标。