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The Protein Journal

, Volume 38, Issue 5, pp 515–524 | Cite as

The Relevance of Glutathione Reductase Inhibition by Fluoxetine to Human Health and Disease: Insights Derived from a Combined Kinetic and Docking Study

  • Ozlem Dalmizrak
  • Kerem Teralı
  • Evelyn Bright Asuquo
  • Izzet Hamdi Ogus
  • Nazmi OzerEmail author
Article

Abstract

Glutathione reductase (GR) is a homodimeric enzyme playing an important role in the regeneration of the central antioxidant molecule reduced glutathione (GSH) from oxidized glutathione (GSSG) at the expense of a molecule of NADPH. GSH scavenges and eliminates superoxide and hydroxyl radicals non-enzymatically or serves as an electron donor for several enzymes. Fluoxetine (FLU), a selective serotonin reuptake inhibitor, is widely prescribed in the treatment of major depressive disorder. Here, using enzyme kinetic studies and molecular docking simulations, we aimed at disclosing the mechanistic and structural aspects of the interaction between GR and FLU. Affecting enzyme activity in a dose-dependent manner, FLU was shown to be a moderately potent (IC50 = 0.88 mM) inhibitor of GR. When the variable substrate was GSSG, the type of inhibition was linear mixed-type competitive (Ki = 279 ± 32 μM; α = 5.48 ± 1.29). When the variable substrate was NADPH, however, the type of inhibition was non-competitive (Ki = 879 ± 82 μM). The observed difference in inhibition types was attributed to the binding of FLU in the large intermonomer cavity of GR, where it hampered catalysis and interfered with substrate binding. Overall, although it is anticipated that long-term use of FLU leads to acquired GR deficiency, the inhibitory action of FLU on GR may be therapeutically exploited in anti-cancer research.

Keywords

Glutathione reductase Fluoxetine Linear mixed-type competitive inhibition Non-competitive inhibition Molecular docking 

Abbreviations

FAD

Flavin adenine dinucleotide

FDA

Food and Drug Administration

FLU

Fluoxetine

GPx

Glutathione peroxidase

GR

Glutathione reductase

GSH

Reduced glutathione

GSSG

Oxidized glutathione, glutathione disulfide

GST

Glutathione S-transferase

IC50

Half-maximal inhibitory concentration

Ki

Inhibition constant, dissociation constant of the enzyme‒inhibitor complex

Km

Michaelis constant

Ks

Dissociation constant of the enzyme‒substrate complex

NADP

Nicotinamide adenine dinucleotide phosphate, oxidized form

NADPH

Nicotinamide adenine dinucleotide phosphate, reduced form

NFLU

Norfluoxetine

PLIP

Protein‒ligand interaction profiler

ROS

Reactive oxygen species

SOD

Superoxide dismutase

SSRI

Selective serotonin reuptake inhibitor

TAS

Total antioxidant status

TPT

Transplacental transfer

Vm

Maximum velocity

Notes

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflicts of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Medical Biochemistry, Faculty of MedicineNear East UniversityMersin 10Turkey

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