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

• 批准号: 10919702
• 负责人: Alexey Zakharov
• 金额: $ 24.08万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 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/10919702
• 关键词: Antibiotic Therapy; Antibiotics; Area; Awareness; Bacteria; Biological Assay; Borrelia; Borrelia burgdorferi; Caseins; Cells; Characteristics; Chronic; Development; Disease; Enzymes; Etiology; Fluorescence Resonance Energy Transfer; Genetic Transcription; Growth; Helicobacter pylori; High temperature of physical object; Homologous Gene; Immune response; Impairment; Label; Literature; Lyme Disease; Molecular Chaperones; Order Spirochaetales; Pathogenesis; Pathogenicity; Patients; Peptides; Post Treatment Lyme Disease Syndrome; Prevalence; Protein Family; Proteins; Quality Control; Recombinants; Reporting; Series; Serine Protease; Specificity; Surface; Testing; Therapeutic Effect; Tick-Borne Diseases; Tissues; Validation; Virulence; antimicrobial; assay development; experience; extracellular; fluorophore; high throughput screening; inhibitor; periplasm; redshift; screening; small molecule; small molecule inhibitor; tool; virtual screening

During the reporting period, the collaborative team began initial assay development during recombinant BbHtrA protein, and enzyme characterization is being completed with a casein derivative labeled with a red-shifted fluorophore. Validation screens with BbHtrA and the casein substrate have demonstrated assay robustness to enable larger-scale screening. 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.

在本报告所述期间，合作小组在重组BbHtrA蛋白期间开始了初步的分析开发，并正在用标记有红移荧光团的酪蛋白衍生物完成酶的鉴定。使用BbHtrA和酪蛋白底物的验证筛选已经证明了能够进行更大规模筛查的检测稳健性。目前正在使用FRET多肽筛选方法对酶的特异性进行进一步的表征，以便获得筛选抑制剂所需的特化FRET底物。