A 4.2 kDa synthetic peptide as a potential probe to evaluate the antibacterial activity of coumarin drugs

✍ Scribed by Andreza C. Scatigno; Saulo S. Garrido; Dr Reinaldo Marchetto

• Book ID: 105360376
• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 161 KB
• Volume: 10
• Category: Article
• ISSN: 1075-2617
• DOI: 10.1002/psc.565

✦ Synopsis

Abstract

The coumarin antibiotics are potent inhibitors of DNA replication whose target is the enzyme DNA gyrase, an ATP‐dependent bacterial type II topoisomerase. The coumarin drugs inhibit gyrase action by competitive binding to the ATP‐binding site of DNA gyrase B protein. The production of new biologically active products has stimulated additional studies on coumarin–gyrase interactions. In this regard, a 4.2 kDa peptide mimic of DNA gyrase B protein from Escherichia coli has been designed and synthesized. The peptide sequence includes the natural fragment 131–146 (coumarin resistance‐determining region) and a segment containing the gyrase–DNA interaction region (positions 753–770). The peptide mimic binds to novobiocin (K~a~ = 1.4 ± 0.3 × 10^5^ M^−1^), plasmid (K~a~ = 1.6 ± 0.5 × 10^6^ M^−1^) and ATP (K~a~ = 1.9 ± 0.4 × 10^3^ M^−1^), results previously found with the intact B protein. On the other hand, the binding to novobiocin was reduced when a mutation of Arg‐136 to Leu‐136 was introduced, a change previously found in the DNA gyrase B protein from several coumarin‐resistant clinical isolates of Escherichia coli. In contrast, the binding to plasmid and to ATP was not altered. These results suggest that synthetic peptides designed in a similar way to that described here could be used as mimics of DNA gyrase in studies which seek a better understanding of the ATP, as well as coumarin, binding to the gyrase and also the mechanism of action of this class of antibacterial drugs. Copyright © 2004 European Peptide Society and John Wiley & Sons, Ltd.