E. coli BW25113 LptH screening strain demonstrates LptH expression when induced with arabinose.
E. coli cells expressing LptH demonstrate lower susceptibility to membrane-targeting antibiotics.
Refolded LptH protein demonstrates a high binding affinity for Lipopolysaccharides.
Deletion of LptA and complementation of LptH in E. coli BW25113 ΔtolC Strain
E. coli contains the protein LptA, a homologue of LptH with a similar function.
Eliminating the effect of LptA will produce more accurate results of the impact of LptH expression on the inhibitory activity of membrane-targeting inhibitors of LPS transport.
Biochemical and cell-based experiments involving LptH
§Use computer-aided virtual screens to identify small molecules predicted to bind LptH.
§Perform binding experiments with purified LptH to determine dissociation constants for selected compounds.
§Use cell-based susceptibility assays to evaluate inhibition of LPS transport.
Employing an ex vivo lung model to test candidate antibiotics against P. aeruginosa
Ex vivo lung tissue models will be infected with P. aeruginosa to test the effectiveness of identified candidate antibiotics to terminate an active infection.