Use of luminescent bacteria for rapid screening and characterization of short cationic antimicrobial peptides synthesized on cellulose using peptide array technology.
The increasing multi-resistance of pathogenic bacteria requires the development of novel classes of antibiotics. Antimicrobial host defense peptides represent one promising class. Here we describe a protocol for screening large numbers of peptides against any microbe of interest. Peptides synthesized on a cellulose support by peptide array technology can be added to a microbe that expresses the luxCDABE (luciferase) gene cassette. Any substance that decreases the energy level within the microbe will cause a quantifiable decrease in light production. The potency of the compound, at different concentrations, is reflected by the rate of decrease in luminescence. In conjunction with peptide array technology, the screening assay is rapid and high throughput and demonstrates good correlation with conventional killing or minimal inhibitory concentration assays performed with the same peptides synthesized by standard solid-phase peptide synthesis. The protocol can be completed in 3 d.
Hilpert
Kai
K
Centre for Microbial Diseases and Immunity Research, University of British Columbia, no. 2259 Lower Mall Research Station, Vancouver, British Columbia, V6T 1Z3, Canada.
Hancock
Robert E W
RE
D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects*; D002412:Cations; D003460:Crystallization; D049449:Luminescence; D008826:Microbial Sensitivity Tests / Q000379:methods; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D040081:Protein Array Analysis / Q000379:methods
D000900:Anti-Bacterial Agents; D002412:Cations; D010455:Peptides