PROTEOMIC PROFILING OF CANDIDA ALBICANS RESPONSES TO NITROSATIVE AND OXIDATIVE

白色念珠菌对亚硝化和氧化反应的蛋白质组学分析

• 批准号: 8167560
• 负责人: DAVID J SINGEL
• 金额: $ 20.02万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167560
• 关键词: Address; Biochemical; Biochemical Pathway; Candida albicans; Cells; Chemical Agents; Chemicals; Chemistry; Color; Computer Retrieval of Information on Scientific Projects Database; Data; Detection; Disulfides; Drug Metabolic Detoxication; Funding; Generations; Goals; Grant; Health; Host Defense; Human; Immune response; Immunocompromised Host; Individual; Institution; Invaded; Label; Measurement; Measures; Methods; Modeling; Modification; Molecular; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Pharmaceutical Preparations; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; Relative (related person); Research; Research Personnel; Resistance; Resources; Risk; Role; Signal Transduction; Source; Stress; Sulfenic Acids; Sulfhydryl Compounds; United States National Institutes of Health; Validation; base; cell growth; computerized data processing; design; macrophage; neutrophil; nitrosative stress; pathogen; programs; response; sensor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Candida albicans is a prevalent human pathogen, which presents a serious health risk in immunocompromised individuals. The first line of host defense against C. albicans involves innate immune responses that challenge the invading pathogen with macrophage- and neutrophil-generated nitrosative and oxidative stress. The counter-measures that pathogens muster against this stress are not well understood, but appear to include roles for proteins involved in enzymatic detoxification of stress-inducing chemical agents, and in signaling cascades that regulate of cell growth. Very little is known, however, at the molecular level, of the responses of C. albicans to oxidative and nitrosative stress. The broad goal of this research is to develop enhanced differential proteomics methods and to apply them to advance our understanding of the molecular bases of C. albicans defenses against host nitrosative and oxidative challenges. Elucidation of the primary biochemical steps in defense pathways will establish new targets for the design of more effective and specific drugs against C. albicans. This research will focus on early responses that are likely to involve reversible modification of reactive protein thiols that yield S-nitrosothiols (RSNO), S- sulfenic acids (RSOH) and transient disulfices. The fundamental questions that we will address are: (1) what components of the C. albicans proteome are modified by different types and levels of oxidative and nitrosative stress; (2) which of these modifications involve redox signaling processes in which protein sensors detect and transduce the stress; and (3) what biochemical pathways are modulated by stress-responsive protein modifications? Differential thiol redox profiling (dThiRP) - featuring a new generation of "Zdyes", that promise unsurpassed threshold sensitivities and the capacity for multi-color differential profiling - will be used to address these questions. Specifically, the research program entails: (1) validation of thiol "switch" detection chemistry with the Zdye labels; (2) global proteomic measurements of changes in protein levels, S-nitrosothiols, sulfenic acids, and disulfides in C. albicans cells subject to exogenous chemical, and macrophage challenge; (3) measurements of the relative resistance to chemical and macrophage challenge of homozygous and heterozygous KO strains, suggested by modeling of the wild-type proteomic data.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 白色念珠菌是一种普遍的人类病原体，在免疫功能低下的个体中呈现出严重的健康风险。对白色念珠菌的宿主防御的第一线涉及先天免疫反应，这些反应挑战了巨噬细胞和中性粒细胞生成的硝酸和氧化应激的入侵病原体。病原体反对这种压力的反测量尚不清楚，但似乎包括参与酶促诱导的化学剂的酶排毒的蛋白质以及调节细胞生长的信号级联反应的作用。然而，在分子水平上，鲜为人知的白色念珠菌对氧化和硝化应激的反应。这项研究的广泛目标是开发增强的差异蛋白质组学方法，并将它们应用它们来促进我们对白色念珠菌防御措施的分子碱基的理解，以防止宿主硝化和氧化挑战。阐明国防途径的主要生化步骤将建立新的目标，以设计针对白色念珠菌的更有效和特定的药物。 这项研究将集中在可能涉及可逆性蛋白硫醇的早期反应上，该硫醇产生S-亚硝基硫醇（RSNO），S-硫酸（RSOH）和瞬时二硫化。我们将要解决的基本问题是：（1）白色念珠菌蛋白质组的哪些组成部分通过不同类型和水平的氧化和硝化应激来修改； （2）这些修饰中的哪一个涉及氧化还原信号传导过程，其中蛋白质传感器检测和转导应力； （3）应力响应蛋白修饰调节了哪些生化途径？ 差异硫醇氧化还原分析（DTHIRP）将使用新一代的“ Zdyes”，有望无与伦比的阈值敏感性和多色差分谱分析的能力 - 将用于解决这些问题。具体而言，研究计划需要：（1）用ZDYE标签验证硫醇“开关”检测化学； （2）在蛋白质水平，S-亚硝基硫醇，硫酸和二硫化物中的全球蛋白质组学测量中，白色念珠菌细胞受到外源性化学化学和巨噬细胞的挑战的变化； （3）测量纯合子和杂合KO菌株对化学和巨噬细胞挑战的相对抵抗力，这是通过对野生型蛋白质组学数据进行建模而提出的。