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

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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.

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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

IC 50 :

Half-maximal inhibitory concentration

K i :

Inhibition constant, dissociation constant of the enzyme‒inhibitor complex

K m :

Michaelis constant

K s :

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

V m :

Maximum velocity

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Authors and Affiliations

  1. Department of Medical Biochemistry, Faculty of Medicine, Near East University, Near East Boulevard, Nicosia/TRNC, Mersin 10, 99138, Turkey

    Ozlem Dalmizrak, Kerem Teralı, Evelyn Bright Asuquo, Izzet Hamdi Ogus & Nazmi Ozer

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  1. Ozlem Dalmizrak
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  2. Kerem Teralı
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  4. Izzet Hamdi Ogus
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Correspondence to Nazmi Ozer.

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Dalmizrak, O., Teralı, K., Asuquo, E.B. et al. The Relevance of Glutathione Reductase Inhibition by Fluoxetine to Human Health and Disease: Insights Derived from a Combined Kinetic and Docking Study. Protein J 38, 515–524 (2019). https://doi.org/10.1007/s10930-019-09834-7

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