Characterization of Disulfide-Bond Formation in Mycobacteria

分枝杆菌中二硫键形成的表征

• 批准号: 8594583
• 负责人: Brian Meehan
• 金额: $ 5.22万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8594583
• 关键词: Adopted; Alkylation; Animal Model; Antibiotic Resistance; Antibiotics; Antibodies; Antitubercular Agents; Bacteria; Biochemical; Biological Assay; Blood coagulation; Cell Wall; Cells; Color; Complement; Cysteine; Databases; Development; Disulfides; Drug resistance in tuberculosis; Electrons; Emergency Situation; Enzymes; Escherichia coli; Extreme drug resistant tuberculosis; Figs - dietary; Galactosidase; Genes; Genetic; Genus Mycobacterium; Growth; Health; Homologous Gene; Human; In Vitro; Integral Membrane Protein; Kinetics; Knowledge; LacZ Genes; Lactose; Mediating; Metabolism; Molecular Conformation; Monitor; Mutation; Mycobacterium smegmatis; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigen 85A; Organism; Oxidants; Pathway interactions; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Play; Process; Protein Disulfide Isomerase; Proteins; Protocols documentation; Quinones; Research; Role; Sequence Homology; System; Testing; Ursidae Family; Virulence; analog; antimicrobial; cell motility; disulfide bond; global health; in vivo; inhibitor/antagonist; mutant; novel; oxidation; pathogen; public health relevance; research study; small molecule; small molecule libraries; success; thioredoxin-like protein; tool

DESCRIPTION (provided by applicant): The rise of extensively drug-resistant Mycobacterium tuberculosis (Mtb) has important implications for human health worldwide. However, the development of new antibiotics directed against this organism has been hindered by a lack of knowledge about many of the processes critical to Mtb replication and virulence. While the genes essential for viability of Mtb have been identified, the physiological roles of many of their products are not well understood. Amongst these vital enzymes are homologs of VKOR and DsbA, proteins that our lab has shown can catalyze the formation of disulfide bonds in proteins secreted by E.coli. Disulfide bonds provide stability to many proteins secreted by bacteria. Chief amongst these are proteins crucial to bacterial cell wall integrity, motility, and virulence. Decades of research focused on the Dsb pathway of E.coli conducted by our lab and others has led to the development of many important genetic and biochemical tools that have provided for a thorough understanding of this system. Experiments proposed here will apply those tools to Mycobacteria so as to characterize what appears to be a novel disulfide bond-forming pathway. The essentiality of VKOR and DsbA suggests that the disulfide bond forming system of Mtb is a highly vulnerable component of metabolism that might be exploited as a target for new antibiotics. Outlined here is a high-throughput approach for identifying small molecule inhibitors of MtbVKOR in the model organism Mycobacterium smegmatis. Using M.smegmatis ¿vkor strains expressing MtbVKOR or E.coli DsbB, we will look for compounds that specifically inhibit VKOR-mediated growth. Such molecules will be important tools for studying disulfide bond formation in vitro, but may also be developed as new antibiotics.

描述（由申请人提供）：广泛耐药结核分枝杆菌（Mtb）的兴起对全球人类健康具有重要影响。然而，由于缺乏对结核分枝杆菌复制和毒力至关重要的许多过程的了解，针对这种生物体的新抗生素的开发受到阻碍。虽然已经确定了对结核分枝杆菌生存能力至关重要的基因，但其中许多基因的生理作用