Bacterial cytological profiling (BCP) is a technique that was developed to rapidly identify not only the bacterial synthesis pathway affected by a given antibiotic, but also the specific target within that pathway. This is accomplished through the use of confocal fluorescence microscopy along with membrane and DNA active fluorescent dyes. It has been proven to work as a method to identify the mechanism of action for novel small molecule antibiotics, reducing the time and number of experiments required to determine how an antibiotic is inhibiting bacterial growth. While this technique has proven effective for small molecule antibiotics, which usually affect only one cellular pathway, it has yet to be determined whether BCP is effective for determining the mechanism of action for antimicrobial peptides. Antimicrobial peptides are capable of affecting more than one area of a cell, and in some cases their mechanisms of action are different from those observed for small molecules. Here we attempt to use BCP to determine the mechanism of action of several antimicrobial peptides. In this work we will perform BCP on several known classes of small molecule antibiotics and well-studied antimicrobial peptides including Magainin 2, Buforin 2, Ixosin, and Piscidins 1 and 3. This work will allow for rapid determination of the mechanism of action for antimicrobial peptides, either eliminating or supplementing the battery of tests currently required to make this determination.

Using bacterial cytological profiling to determine the mechanism of action of antimicrobial peptides

256th National Meeting (2018)

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