Biochemical and biophysical characterization of a mycoredoxin protein glutaredoxin A1 from Corynebacterium pseudotuberculosis

Autor(es)

Eberle, Raphael J. ; Kawai, Liege A. ; de Moraes, Fabio R. ; Tasic, Ljubica ; Arni, Raghuvir K. ; Coronado, Monika A.

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 107, p. 9-pg., 2018-02-01.

Resumo

Glutaredoxin A1 from Corynebacterium pseudotuberculosis was shown to be a mycoredoxin protein. In this study, we established a process to overexpress and purify glutaredoxin A1. The aim of this study was the investigation of the Glutaredoxin A1 from C pseudotuberculosis behavior under different redox environments and the identification of lead molecules, which can be used for specific inhibitor development for this protein family. A quantitative assay was performed measuring the rate of insulin reduction spectrophotometrically at 640 nm through turbidity formation from the precipitation of the free insulin. Glutaredoxin A1, at 5 mu M concentration, accelerated the reduction process of 0.2 mM insulin and 1 mM DTT. The pH optimum of the reaction was 7.4. In the presence of DTT and ESH the glutaredoxin A1 presents similar activity, and its activity is reduced by 50% in the presence of GSH. Additional function for ESH in the redox metabolism of C. pseudotuberculosis is suggested. A combined STD and Chemical Shift - NMR approach was employed to study the effects of potential inhibitors on the structure of glutaredoxin A1 from Corynebacterium pseudotuberculosis. The inhibitory potential of four ligands (heparin, suramin, hesperetin - Hst, and hesperidin - Hsp) against glutaredoxin A1 is discussed. (C) 2017 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 09/53989-4 - EMU: aquisição de espectrômetro de ressonância magnética nuclear para estudos de biomoléculas
• Beneficiário: Raghuvir Krishnaswamy Arni
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 15/13765-0 - ESTUDOS ESTRUTURAIS E CARACTERIZAÇÃO DE PROTEÍNAS POR CRISTALOGRAFIA DE RAIOS-X E RESSONÂNCIA MAGNETICA NUCLEAR. Investigações estruturais e biofísicas dos mecanismos moleculares de proteínas funcionais
• Beneficiário: Raghuvir Krishnaswamy Arni
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/08104-8 - Aspectos funcionais e estruturais de duas proteínas de ligação de DNA codificadas pela Corynebacterium pseudotuberculosis
• Beneficiário: Raphael Josef Eberle
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/18868-2 - Aquisição de equipamento multiusuário para biologia molecular e estrutural
• Beneficiário: Raghuvir Krishnaswamy Arni
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 16/12904-0 - Mecanismos e Interações Moleculares de Moléculas Bioativas com a Protease NS3 do Zika Vírus.
• Beneficiário: Monika Aparecida Coronado
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado