Targeting aphA: A new high throughput screening assay identifies compounds that reduce prime virulence factors of Vibrio cholerae.

Targeting aphA: A new high throughput screening assay identifies compounds that reduce prime virulence factors of Vibrio cholerae. J Med Microbiol. 2016 May 12; Authors: Bolger G, Roy S, Zapol'skii VA, Kaufmann DE, Schnürch M, Mihovilovic MD, Nandy RK, Tegge W Abstract A high throughput screening (HTS) assay was developed for identifying compounds with inhibitory effect on aphA, one of the key regulators positively controlling Vibrio cholerae pathogenesis. An inhibitory effect on aphA was expected to lead to attenuation in the secretion of the major pathogenicity factors of V. cholerae, cholera toxin (CT) and toxin co-regulated pilus (TCP). The plasmid construct pAKSB was developed with a kanamycin resistance (KmR) gene under the control of the aphA like promoter for conferring KmR phenotype under aphA expressing conditions. The HTS assay was performed to identify compounds with inhibitory effect on the growth of O139 V. cholerae MO10 carrying the construct pAKSB in growth medium containing Km (30 g ml-1), but not in its absence. Of 20,338 compounds screened, six compounds were identified to inhibit the pAKSB induced KmR phenotype and these compounds caused transcriptional inhibition of aphA in V. cholerae O139 strain MO10 as well as variant V. cholerae O1 El Tor strain NM06-058. Of the three most active substances, compound 53760866 showed lowest half maximal cytotoxicity (c-IC50) in a eukaryotic cell viability assay and was cha...
Source: Journal of Medical Microbiology - Category: Microbiology Authors: Tags: J Med Microbiol Source Type: research