title: Fluorescence-based methods to assay inhibitors of lipopolysaccharide synthesis.
abstract: Treatment of infections caused by Gram-negative bacteria is difficult due in large part to problems arising from innate and acquired drug resistance, resulting in a limited number of effective antibiotics. Consequently, antibiotics that can circumvent mechanisms of drug resistance are needed. Lipid A is a glucosamine phospholipid that acts as an anchor for lipopolysaccharides (LPS) that comprise the outer membranes of Gram-negative bacteria, a barrier for small molecule entry into the cell, and is also the portion of LPS that stimulates the immune system in septic shock. Consequently, inhibitors of lipid A biosynthesis have the potential to function as antibiotics and/or anti-endotoxins in the treatment of Gram-negative bacterial infections. Current efforts in the development of antibiotics targeted against lipid A have focused on the metal-dependent deacetylase LpxC. Herein we describe fluorescence-based assays that can be used for the evaluation of LpxC inhibitors with the potential to serve as antibiotics.
authors.affiliation: Department of Biochemistry, Virginia Tech, Blacksburg, VA, USA. hernickm@vt.edu
authors.forename: Marcy
authors.initials: M
authors.lastname: Hernick
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D005453:Fluorescence; D005454:Fluorescence Polarization / Q000379:methods*; D006090:Gram-Negative Bacteria / Q000187:drug effects; D008050:Lipid A / Q000096:biosynthesis*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D008050:Lipid A; D000581:Amidohydrolases; C097307:UDP-3-O-acyl-N-acetylglucosamine deacetylase