Novel in silico to in vitro schema: new antibiotic exploration approach

Docking prediction analysis is a computerized model which can aid in developing new potential bacterial targets for antibiotics. Trying to find a molecule with a high potential binding with its macromolecule (protein) is challenging. The bacterial cross talk (quorum sensing [QS]) mechanisms are bacterial regulation processes which allow bacteria to express pathogenic phenotypes such as biofilm formation. By targeting the QS components, a novel way of affecting bacterial behavior is achieved, possibly without creating antibiotic resistance. We conducted a combined analysis of a new synthetic drug with a docking in silico prediction applied on potential QS targets in Pseudomonas aeruginosa , and by doing so validated previous docking results of other QS inhibitors. We complemented the in silico analysis with an in vitro E. coli - QS reporting assay in order to confirm a new class of QS inhibitors. Our analysis was confirmed after conducting site-directed mutagenesis over the sequences of the potential binding amino acid residues of the suspected QS inhibitor target. Our analysis was supported by a wide meta-genomic microarray analysis which revalidated the QS inhibition schema. The QS inhibitor successfully exhibited a decrease in the pathogenic phenotypes of surface attached biofilm and swarming motility in P. aeruginosa .