Abstract: Lipopolysaccharides are an essential component of the membrane of gram-negative bacteria, which cause a multitude of diseases such as cholera and influenza. Inhibiting lipopolysaccharide production by targeting a key enzyme in its synthetic pathway, Heptosyltransferase I, has been shown to increase bacterial susceptibility to antibiotics. Previously in the Taylor lab, 419 compounds were computationally docked onto HepI using Autodock Vina. The molecules predicted to have the worst and best binding energies were tested in inhibition assays; the resulting inhibition constants did not match the predictions, implying that the data has to be analysed more carefully. This summer, we successfully extracted the substrates of HepI, namely ADP-Heptose and O-deacylated Lipid A. HepI protein was also expressed and purified. An ADP/NADH coupled kinetics assay was then performed with varying ODLA concentrations to perfect the technique of measuring enzymatic rate. Our current objective is to perform statistical analysis to find possible correlations between the computational and experimental data that correctly predict the observed inhibition trends. The updated hypotheses will then be tested using the same kinetics assay to identify potent HepI inhibitors to be used in drug development.
Live Poster Session:
Thursday, July 29th 1:15-2:30pm EDT