Oxygen Sensors and P450 Monooxygenases in Mycobacertium tuberculosis

结核分枝杆菌中的氧传感器和 P450 单加氧酶

基本信息

批准号：
7294665
负责人：
Paul R Ortiz De Montellano
金额：
$ 40.69万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7294665
关键词：
Acquired Immunodeficiency Syndrome Aerobic Affinity Antitubercular Agents Area Attenuated Azoles Bacillus (bacterium)Binding Biochemical Biological Cessation of life Clinic Commercial Sectors Complement Condition Cytochrome P450 DNA Microarray Chip DNA Microarray format Development Disease Disruption Drug Delivery Systems Drug Design Drug resistance Enzymes Evaluation Family Gases Genes Genome Genus Mycobacterium Goals Growth Hand Health Heel Heme Heme Group Heme Iron Hemeproteins Histidine In Vitro Incidence Individual International Knock-out Label Laboratories Lead Length Libraries Ligands Link Literature Methods Mixed Function Oxygenases Modeling Mus Mycobacterium tuberculosis Organism Outcome Oxygen Persons Pharmaceutical Preparations Phosphotransferases Physiological Population Prevalence Public Health Publishing Regulon Relative (related person)Research Research Personnel Resistance Resistance development Role Route Scourge Screening procedure Sensory Signal Transduction Site Sterilization for infection control Structure System Techniques Testing Therapeutic Therapeutic Agents Thinking Translating Tuberculosis United States United States National Institutes of Health Update Virulence Work base cost drug development experience fungus high throughput screening in vivo inhibitor/antagonist interest latent infection member men mortality mycobacterial programs protein-histidine kinase receptor sensor synergism tuberculosis drugs tuberculosis treatment

Acquired Immunodeficiency Syndrome Aerobic Affinity Antitubercular Agents Area Attenuated Azoles Bacillus (bacterium)Binding Biochemical Biological Cessation of life Clinic Commercial Sectors Complement Condition Cytochrome P450 DNA Microarray Chip DNA Microarray format Development Disease Disruption Drug Delivery Systems Drug Design Drug resistance Enzymes Evaluation Family Gases Genes Genome Genus Mycobacterium Goals Growth Hand Health Heel Heme Heme Group Heme Iron Hemeproteins Histidine In Vitro Incidence Individual International Knock-out Label Laboratories Lead Length Libraries Ligands Link Literature Methods Mixed Function Oxygenases Modeling Mus Mycobacterium tuberculosis Organism Outcome Oxygen Persons Pharmaceutical Preparations Phosphotransferases Physiological Population Prevalence Public Health Publishing Regulon Relative (related person)Research Research Personnel Resistance Resistance development Role Route Scourge Screening procedure Sensory Signal Transduction Site Sterilization for infection control Structure System Techniques Testing Therapeutic Therapeutic Agents Thinking Translating Tuberculosis United States United States National Institutes of Health Update Virulence Work base cost drug development experience fungus high throughput screening in vivo inhibitor/antagonist interest latent infection member men mortality mycobacterial programs protein-histidine kinase receptor sensor synergism tuberculosis drugs tuberculosis treatment

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项目摘要

DESCRIPTION (provided by applicant): The three greatest current needs for progress in the treatment of tuberculosis are to (a) develop drugs against drug resistant strains of Mycobacterium tuberculosis, (b) develop effective approaches to treat the latent states of the disease, and (c) shorten the course of therapy, which is related to the second goal. Here we propose to characterize the DevS/DosR and the redundant DosT/DosR two-component regulatory systems of M. tuberculosis. These regulatory systems control a panel of approximately 50 genes that is induced by low oxygen concentrations or NO exposure. The anaerobic state of M. tuberculosis is thought to resemble (but not be identical to) that found in latent infections and serves as model for it. We have found that the DevS and DosT sensor kinases have a heme group in their sensory domain and will investigate the structure of these sensors, their sensitivity to O2, CO, and NO, and the mechanisms that link binding of a gas to the heme group with autophosphorylation of a histidine residue in the kinase. A high-throughput screen will be performed to identify inhibitors of the DevS / DosT kinases, as inhibitors offer a possible route to treatment of the latent disease. Determination of structure-activity profiles and optimization of the inhibitors will be pursued. In a related but distinct effort, we propose to clone, express, and characterize the P450 enzyme complement of M. tuberculosis. Clinically used azole drugs that target P450 enzymes in fungi attenuate mycobacterial growth even in the latent state, but the substrates and biological roles of the twenty M. tuberculosis P450 enzymes are not known. Two of the P450 enzymes have been described in the literature, and we already have three further members of this M. tuberculosis family in hand. Each enzyme will be characterized by spectroscopic and crystallographic methods, and its affinity for azole drugs will be determined. A search for the relevant substrates for each enzyme will be pursued using a variety of techniques. The role of the P450 enzymes will also be investigated by individually disrupting ("knocking out") each P450 enzyme in M. tuberculosis. The consequences of each P450 enzyme disruption will be analyzed by DNA microarray studies in addition to determination of the associated changes in bacterial growth and virulence. The project thus encompasses two heme- and oxygen-dependent system of M. tuberculosis that offer new avenues to the development of drugs effective against the latent form of the disease.

描述（由申请人提供）：结核病治疗的三个最大目前需求是（a）开发针对结核分枝杆菌抗药性菌株的药物，（b）开发有效治疗疾病潜伏状态的有效方法，以及（c）缩短治疗过程，这与第二个目标有关。在这里，我们建议表征Devs/DOSR和结核分枝杆菌的冗余DOST/DOSR两组调节系统。这些调节系统控制着大约50个基因的面板，该面板由低氧浓度或没有暴露诱导。结核分枝杆菌的厌氧状态被认为与潜在感染中发现的（但不相同）并作为其模型。我们发现，DEV和DOST传感器激酶在其感觉域中具有血红素组，并将研究这些传感器的结构，对O2，CO和NO的敏感性以及将气体与血红素组结合与血红素基团与激酶中组氨酸残基的自磷酸化的机制。将进行高通量屏幕以鉴定Devs / DOST激酶的抑制剂，因为抑制剂提供了治疗潜在疾病的可能途径。确定结构活性概况和抑制剂的优化。在相关但独特的努力中，我们提议克隆，表达和表征结核分枝杆菌的P450酶补体。临床上使用的偶氮药物在真菌中靶向P450酶，即使在潜在状态下也会减弱分枝杆菌的生长，但是二十种结核分枝杆菌P450酶的底物和生物学作用却尚不清楚。文献中已经描述了P450酶中的两种，我们手头上还有三个成员。每种酶将以光谱和晶体学方法的特征，并确定其对硫唑药物的亲和力。搜索每个酶的相关底物将使用各种技术进行追求。 P450酶的作用也将通过单独破坏（“击倒”）在结核分枝杆菌中的每个P450酶来研究。除了确定细菌生长和毒力的相关变化外，DNA微阵列研究还将分析每种P450酶破坏的后果。因此，该项目涵盖了两个血红素和氧依赖性结核分枝杆菌的系统，这些系统为有效的药物开发了针对该疾病的潜在形式的新途径。