Principles of Protein Mimicry of DNA

DNA 蛋白质模拟原理

基本信息

批准号：
7758713
负责人：
Michael D. Brenowitz
金额：
$ 33.1万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-20 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7758713
关键词：
Antibiotic Resistance Antibiotics Bacteria Binding Binding Proteins Biochemical Biological Biological Models Biological Process Characteristics Charge Clinical Complex DNA DNA Gyrase DNA Mimicry DNA analysis Development Drug resistance Electrostatics Enzymes Family Fluoroquinolones Foundations Genus Mycobacterium Goals Hand Heart Human Investigation Ions Laboratories Mediating Metabolic Pathway Modeling Molecular Mutagenesis Mycobacterium tuberculosis Nucleic Acids Peptides Plasmids Play Process Property Protein Family Protein Footprinting Proteins Quinolones Regulation Resistance Resolution Role Scaffolding Protein Shapes Specificity Structure Surface Testing Therapeutic Therapeutic Index Thermodynamics Toxic effect Tuberculosis Vision Xanthomonas albilineans base driving force fluoroquinolone resistance member mimicry new therapeutic target novel pathogen physical property planetary Atmosphere protein folding protein function therapeutic protein

项目摘要

DESCRIPTION (provided by applicant): This revised application of a new proposal describes a comprehensive biochemical and biophysical investigation into the `DNA mimicry' of a recently discovered member of the Pentapeptide Repeat Protein (PRP) family of proteins. The atomic resolution structure of the MfpA PRP from Mycobacterium tuberculosis revealed a new protein fold; MfpA's right-handed 2-helical coils yield an elongated structure that is a remarkable mimic of the size, shape and electrostatic surface of DNA. That DNA mimicry is important to the biological role of MfpA is suggested by its ability to confer resistance to fluoro- quinolones by binding DNA gyrase thereby inhibiting its function. We will test the hypothesis that mimicry of the physical properties DNA by MfpA is the heart of its ability to confer drug resistance. Our long term goal is to understand the molecular mechanism by which MfpA and other PRP mediate antibiotic resistance and potentially regulate the enzymatic processing of DNA. MfpA will be developed as a model system for the exploration of DNA mimicry by this newly-discovered and widely-distributed class of proteins. Fluoroquinolones are extremely important antibiotics due to their efficacy against Gram- positive, Gram-negative and mycobacteria. Fluoroquinolones act on DNA gyrase with a unique mechanism of action. They are remarkable for their absence of toxicity and a therapeutic index that is the highest of all currently prescribed antibiotics. The rapid emergence, and spread, of transmissible forms of fluoroquinolone resistance due to the expression of the plasmid-encoded proteins threatens the clinical utility of these antibiotics. The proposed studies seek to understand how the physical properties of the Peptapeptide Repeat Proteins MfpA and Qnr generate resistance to fluoroquinolones. This understanding would provide new targets for therapeutics complementary to fluoroquinolone that would counter the PRP-mediated resistance. Conversely, an understanding of the mechanism by which these proteins function may allow the PRP fold to be used as a platform for the development of novel protein or peptide therapeutics.

描述（由申请人提供）：这项新提案的修订应用描述了对最近发现的五肽重复蛋白（PRP）蛋白质家族的“ DNA模仿”的全面生化和生物物理研究。结核分枝杆菌MFPA PRP的原子分辨率结构显示出了新的蛋白质折叠。 MFPA的右手2螺旋线圈产生的细长结构，是DNA的大小，形状和静电表面的显着模仿。 DNA模拟对MFPA的生物学作用很重要，它是由于其通过结合DNA陀螺酶赋予氟喹诺酮类抗性的能力而抑制其功能。我们将检验以下假设：MFPA模仿物理性质DNA是其赋予耐药性能力的核心。我们的长期目标是了解MFPA和其他PRP介导抗生素耐药性并可能调节DNA的酶促加工的分子机制。 MFPA将作为模型系统开发，用于探索这种新发现且分布广泛的蛋白质类别的DNA模拟。氟喹诺酮类药物是极其重要的抗生素，因为它们对革兰氏阴性，革兰氏阴性菌和分枝杆菌有效。氟喹诺酮类作用于DNA回旋酶具有独特的作用机理。它们因缺乏毒性和当前处方抗生素中最高的治疗指数而着称。由于质粒编码的蛋白的表达，氟喹诺酮抗性的可传播形式的快速出现和传播威胁着这些抗生素的临床实用性。拟议的研究试图了解肽肽重复蛋白MFPA和QNR的物理特性如何产生对氟喹诺酮类的抗性。这种理解将为氟喹诺酮辅助治疗提供新的靶标，这将抵抗PRP介导的抗性。相反，对这些蛋白质功能可能允许PRP折叠的机制的理解可以用作开发新型蛋白质或肽疗法的平台。