Screening for small molecule inhibitors of B. burgdorferi HtrA

这是马里兰大学的Utpal Pal博士的实验室和NCATS的ETB共同努力的一个初步项目，通过NIH-GPP博士前个人伙伴计划得到推动。 重组BbHtrA蛋白已经转移到初步的检测开发中，用红移荧光团标记的酪蛋白衍生物正在完成酶的表征。使用BbHtrA和使用脱机Lopac文库的酪蛋白底物的验证筛选表明，该检测在重复性和稳健性方面具有可接受的统计数据，正在进行进一步的测试，以将该检测带入在线系统。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。

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.