Screening for small molecule inhibitors of B. burgdorferi HtrA

National Center For Advancing Translational Sciences

Investigators

Abstract

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.