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Stabilization and regulation of a Bacillus anthracis spore lytic enzyme

炭疽芽孢杆菌孢子裂解酶的稳定和调节

基本信息

批准号：
8623183
负责人：
DAVID L POPHAM
金额：
$ 22.36万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-12-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8623183
关键词：
Address Bacillus anthracis Bacillus anthracis spore Bacteria Bacterial Spores Biochemical Biochemical Genetics Biological Assay Bypass Chemicals Chimeric Proteins Commit Communicable Diseases Complement Complex Data Decontamination Disease Effectiveness Environment Enzymes Event Food Genes Genetic Genetic Screening Germination Goals Homology Modeling Hydrolysis Immune Sera Industrial Product Infectious Agent Knowledge Left LytA enzyme Lytic Methods Modeling Mutagenesis Mutation Organism Peptide Hydrolases Peptides Peptidoglycan Pharmaceutical Preparations Pharmacologic Substance Play Poisoning Population Process Production Property Protein Region Proteins Regulation Reproduction spores Resistance Role Signal Pathway Site Structure Technology Vaccines Variant Work X-Ray Crystallography antimicrobial cell type crosslink his6 tag improved in vivo innovation killings prevent protein complex protein function public health relevance structural biology

项目摘要

Elimination of spores of Bacillus anthracis and other disease-causing bacteria from contaminated sites or materials is challenging due to the extreme resistance of spores to conventional antimicrobial treatments. Activation of the B. anthracis spore cortex peptidoglycan lytic machinery results in rapid loss of spore dormancy and resistance properties. This machinery can thus be targeted for germination-activating and spore-killing treatments. The cortex lytic enzyme SleB is stored in an inactive and highly stable form in the dormant spore and becomes active early during the germination process. The YpeB protein is required for incorporation of SleB into the dormant spore, and during germination, proteolytic cleavage of YpeB releases SleB to depolymerize the cortex. Aim 1 of the proposed work includes biochemical, genetic, and structural biology methods to reveal protein-protein contacts formed by YpeB and SleB in the dormant spore. Protein complexes containing YpeB or SleB will be purified from spores and co-purified proteins will be identified. Altered forms of YpeB will be expressed in vivo to identify key regions of the protein involved in function and protein interaction. The YpeB structure will be determined using X-ray crystallography in order to understand the effects of mutations. This work will clarify the mechanism of SleB stabilization, which may be adaptable to the production of stable protein pharmaceuticals, vaccines, and other industrial products. Aim 2 is the identification of the protease that processes YpeB and thus activates SleB. Knowledge of this protease and its regulation will suggest methods for the activation of germination via YpeB and SleB. The proposed work will contribute to two major translational goals: Activation of spore germination in order to improve decontamination and understanding of a mechanism of extreme protein stabilization.

消除炭疽芽孢杆菌和其他致病细菌的孢子