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

Author(s)

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

• Document type: Journal article
• Source: International Journal of Biological Macromolecules; v. 107, p. 9-pg., 2018-02-01.

Abstract

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)

• FAPESP's process: 09/53989-4 - Acquisition of a nuclear magnetic resonance spectrometer for studies of biomolecules
• Grantee: Raghuvir Krishnaswamy Arni
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 15/13765-0 - Structural Studies and Characterization of Proteins by X-ray Crystallography and Nuclear Magnetic Resonance. Structural investigations and biophysics of molecular mechanisms of functional proteins.
• Grantee: Raghuvir Krishnaswamy Arni
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/08104-8 - Structural and functional aspects of two DNA binding proteins encoded by Corynebacterium pseudotuberculosis
• Grantee: Raphael Josef Eberle
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/18868-2 - Multi-user equipment acquisition for molecular and structural biology
• Grantee: Raghuvir Krishnaswamy Arni
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 16/12904-0 - Mechanism and Molecular Interactions of Bioactive molecules with NS3 protease from Zika virus.
• Grantee: Monika Aparecida Coronado
• Support Opportunities: Scholarships in Brazil - Post-Doctoral