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Screening for small molecule inhibitors of B. burgdorferi HtrA

伯氏疏螺旋体HtrA小分子抑制剂的筛选

基本信息

批准号：
10691142
负责人：
Alexey Zakharov
金额：
$ 25.96万
依托单位：
NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10691142
关键词：
Aftercare Antibiotic Therapy Antibiotics Area Awareness Bacteria Biological Assay Borrelia Borrelia burgdorferi Caseins Cells Centers for Disease Control and Prevention (U.S.)Characteristics Chronic Development Disease Enzymes Etiology Fluorescence Resonance Energy Transfer Genetic Transcription Growth Helicobacter pylori High temperature of physical object Immune response Impairment Individual Label Libraries Literature Lyme Disease Maryland Molecular Chaperones National Center for Advancing Translational Sciences Online Systems Order Spirochaetales Pathogenesis Pathogenicity Patients Peptides Prevalence Protein Family Proteins Quality Control Recombinants Reproducibility Series Serine Protease Specificity Surface Syndrome Testing Therapeutic Effect Tick-Borne Diseases Tissues United States National Institutes of Health Universities Validation Virulence antimicrobial assay development experience extracellular fluorophore high throughput screening inhibitor periplasm pre-doctoral programs screening small molecule small molecule inhibitor statistics tool virtual screening

项目摘要

This is an incipient project as a combined effort between Dr. Utpal Pal's lab at the University of Maryland and ETB at NCATS, facilitated through an NIH-GPP Pre-Doctoral Individual Partnership Program. Recombinant BbHtrA protein has been transferred for initial assay development, and enzyme characterization is being completed with a casein derivative labelled with a red-shifted fluorophore. Validation screens with BbHtrA and the casein substrate using offline LOPAC libraries show that the assay has acceptable statistics for reproducibility and robustness, and further testing is being conducted to bring the assay to online systems. Further characterization of enzyme specificity is being conducted using a FRET peptide screening approach in order to move towards a characterized FRET substrate necessary for screening for inhibitors.

这是一个早期的项目，作为Utpal博士在马里兰州大学的实验室和NCATS的ETB之间的联合努力，通过NIH-GPP博士前个人合作计划提供便利。重组BbHtrA蛋白已被转移用于初步检测开发，酶的表征正在用红移荧光团标记的酪蛋白衍生物完成。使用离线LOPAC文库对BbHtrA和酪蛋白底物进行的验证筛选显示，该测定具有可接受的重现性和耐用性统计数据，正在进行进一步测试以将该测定引入在线系统。酶特异性的进一步表征正在使用FRET肽筛选方法进行，以便向筛选抑制剂所需的表征FRET底物移动。