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.

这是一个初期的项目，作为马里兰州大学的UTPAL PAL博士实验室与NCAT的ETB之间的共同努力，并通过NIH-GPP博士前的个人合作伙伴计划促进。重组BBHTRA蛋白已转移以进行初始测定开发，并使用带有红移的荧光团标记的酪蛋白衍生物完成酶的表征。使用离线LOPAC库的BBHTRA和酪蛋白基材的验证屏幕表明，该测定法具有可接受的可重复性和鲁棒性的可接受统计信息，并且正在进行进一步的测试，以将测定方法带入在线系统。正在使用FRET肽筛选方法进行酶特异性的进一步表征，以便朝着抑制剂筛选所需的特征FRET底物移动。