Probing the binding of the coumarin drugs using peptide fragments of DNA gyrase B protein

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorGarrido, Saulo Santesso
dc.creatorScatigno, A. C.
dc.creatorTrovatti, E.
dc.creatorCarvalho, D. C.
dc.creatorMarchetto, Reinaldo
dc.date2014-05-27T11:21:19Z
dc.date2016-10-25T18:20:42Z
dc.date2014-05-27T11:21:19Z
dc.date2016-10-25T18:20:42Z
dc.date2005-05-01
dc.date.accessioned2017-04-06T01:13:02Z
dc.date.available2017-04-06T01:13:02Z
dc.identifierJournal of Peptide Research, v. 65, n. 5, p. 502-511, 2005.
dc.identifier1397-002X
dc.identifierhttp://hdl.handle.net/11449/68214
dc.identifierhttp://acervodigital.unesp.br/handle/11449/68214
dc.identifier10.1111/j.1399-3011.2005.00264.x
dc.identifierWOS:000229115800004
dc.identifier2-s2.0-18344372591
dc.identifierhttp://dx.doi.org/10.1111/j.1399-3011.2005.00264.x
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/889581
dc.descriptionBacterial DNA gyrase, has been identified as the target of several antibacterial agents, including the coumarin drugs. The coumarins inhibit the gyrase action by competitive binding to the ATP-binding site of DNA gyrase B (GyrB) protein. The high in vitro inhibitory potency of coumarins against DNA gyrase reactions has raised interest in studies on coumarin-gyrase interactions. In this context, a series of low-molecular weight peptides, including the coumarin resistance-determining region of subunit B of Escherichia coli gyrase, has been designed and synthesized. The first peptide model was built using the natural fragment 131-146 of GyrB and was able to bind to novobiocin (K a = 1.8 ± 0.2 × 105/M) and ATP (Ka = 1.9 ± 0.4 × 103/M). To build the other sequences, changes in the Arg136 residue were introduced so that the binding to the drug was progressively reduced with the hydrophobicity of this residue (Ka = 1.3 ± 0.1 × 105/M and 1.0 ± 0.2 × 105/M for Ser and His, respectively). No binding was observed for the change Arg136 to Leu. In contrast, the binding to ATP was not altered, independently of the changes promoted. On the contrary, for peptide-coumarin and peptide-ATP complexes, Mg2+ appears to modulate the binding process. Our results demonstrate the crucial role of Arg 136 residue for the stability of coumarin-gyrase complex as well as suggest a different binding site for ATP and in both cases the interactions are mediated by magnesium ions. Copyright Blackwell Munksgaard, 2005.
dc.languageeng
dc.relationJournal of Peptide Research
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectAffinity chromatography
dc.subjectCoumarins
dc.subjectDNA gyrase
dc.subjectFluorescence
dc.subjectPeptide synthesis
dc.subjectQuenching
dc.subjectcoumarin anticoagulant
dc.subjectDNA topoisomerase (ATP hydrolysing) B
dc.subjectnovobiocin
dc.subjectaffinity chromatography
dc.subjectbinding site
dc.subjectcomplex formation
dc.subjectDNA supercoiling
dc.subjectdrug binding
dc.subjectenzyme activity
dc.subjectenzyme subunit
dc.subjectnonphotochemical quenching
dc.subjectpeptide synthesis
dc.subjectpriority journal
dc.subjectprotein binding
dc.subjecttarget cell
dc.subjectAdenosine Triphosphate
dc.subjectAmino Acid Sequence
dc.subjectArginine
dc.subjectBinding Sites
dc.subjectBinding, Competitive
dc.subjectChromatography, Affinity
dc.subjectDNA Gyrase
dc.subjectDrug Design
dc.subjectEscherichia coli Proteins
dc.subjectMagnesium
dc.subjectMolecular Sequence Data
dc.subjectNovobiocin
dc.subjectPeptide Fragments
dc.subjectBacteria (microorganisms)
dc.subjectEscherichia coli
dc.titleProbing the binding of the coumarin drugs using peptide fragments of DNA gyrase B protein
dc.typeOtro


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