Ferrand, Sandrine, Tao, Jianshi, Shen, Xiaoyu, Mcguire, Dorothy, Schmid, Andres, Glickman, Fraser and Schopfer, Ulrich (2011) Screening for Mevalonate Pathway Inhibitors using sensitized bacteria strains. Journal of Biomolecular Screening, 16 (6). pp. 637-646. ISSN 1087-0571

Abstract

A simple, optical density based assay for mevalonate pathway inhibitors was developed, that relies on sensitized S. aureus strains and that is fully compatible with high-density screening in 1536-well format. S. aureus strains were constructed in which the mevalonate pathway genes are regulated by an IPTG (isopropyl--D-thiogalactopyranoside)-inducible promoter. Inhibitors of the target enzyme appeared more potent under low IPTG conditions, where a lower amount of enzyme is available to the bacteria, than under high IPTG conditions. This effect was used to bias the hits towards specific inhibitors. Screens were run against the enzymes HMG-CoA synthase (MvaS) and mevalonate kinase (mvaK1), mevalonate diphosphate decarboxylase (mvaD) and phosphomevalonate kinase (mvaK2). The later three enzymes are regulated as an operon. These assays allowed the discovery of potent antibacterial leads that led to an active lead optimization program. The example presented here demonstrates that a sensitization strategy can be successfully applied to > 1 Mio compound HTS in a high density 1536-well format.

Item Type:	Article
Keywords:	Mevalonate pathway, Staphylococcus aureus, sensitized strain, optical density, High Throughput Screening, MRSA, High Throughput Cherry Picking, Infectious disease.
Date Deposited:	29 Jul 2016 00:45
Last Modified:	29 Jul 2016 00:45
URI:	https://oak.novartis.com/id/eprint/3775