Purification of Glutathione Reductase from Human Erythrocytes: Inhibition Profile of Some Anti-Epileptic Drugs

Yıl 2019, , 2140 - 2147, 01.12.2019

Yeliz Demir

https://doi.org/10.21597/jist.525154

Cited By: 6

Öz

Glutathione reductase (GR) is found in the NADPH-dependent oxidoreductase family. GR has various important functions in the cell, such as protein and DNA biosynthesis, the detoxification of reactive oxygen species and free radicals. The purpose of this research was to perform the in vitro inhibition effects of anti-epileptic drugs (phenytoin, gabapentin, and primidone) on GR enzyme. In the current study, the GR enzyme was purified from human erythrocytes with a specific activity of 20.08 EU/mg protein and 2135.97-purification fold. To determine the inhibition effects of anti-epileptic drugs on GR enzyme, Lineweaver-Burk graphs were drawn for each inhibitor. Ki values and inhibition types were determined from these plotted graphs. The Ki values of drugs were found in ranging from 0.15± 0.03-5.74±1.14 mM. Phenytoin was shown the most effective inhibitor feature with a competitive inhibition type.

Anahtar Kelimeler

anti-epileptic drugs, enzyme inhibition, enzyme purification, glutathione reductase

Glutatyon Redüktaz Enziminin İnsan Eritrositlerinden Saflaştırılması: Bazı Anti-epileptik ilaçların İnhibisyon Profili

Öz

Glutatyon redüktaz (GR), NADPH'ye bağlı oksidoredüktaz ailesinde bulunur. GR hücrede protein ve DNA biyosentezi, reaktif oksijen türlerinin ve serbest radikallerin detoksifikasyonu gibi çeşitli önemli fonksiyonlara sahiptir. Bu çalışmanın amacı, GR enzimi üzerine antiepileptik ilaçların (fenintoin, gabapentin ve pirimidon) in vitro etkilerini belirlemektir. Bu çalışmada, GR enzimi insan eritrositlerinden 20.08 EU/mg protein spesifik aktivite ve 2135.97 kat saflaştırıldı. Anti-epileptik ilaçların GR enzimi üzerindeki inhibisyon etkisini belirlemek için Lineweaver-Burk grafikleri çizildi; Ki sabiti ve inhibisyon tipleri bu çizilen grafiklerden hesaplandı. Ki değerleri 0.15± 0.03-5.74±1.14 mM aralığında bulundu. Fenintoin en etkili inhibitör özelliğini yarışmalı inhibisyon tipi ile göstermiştir.

Anahtar Kelimeler

Anti-epileptik ilaçlar, enzim inhibisyonu, enzim saflaştırılması, glutatyon redüktaz

Kaynakça

• Akkemik E, Şenturk M, Özgeriş FB, Taşer, P, Çiftci M, 2011. In vitro effects of some drugs on human erythrocyte glutathione reductase. Turkish Journal of Medical Sciences, 41(2): 235-241.
• Alım Z, Kılıc D, Demir Y, 2018. Some indazoles reduced the activity of human serum paraoxonase 1, an antioxidant enzyme: in vitro inhibition and molecular modeling studies. Archives of Physiology and Biochemistry, 125(5):387-395
• Aslan HE, Demir Y, Ozaslan MS, Türkan F, Beydemir S, Kufrevioglu OI, 2018. The behavior of some chalcones on acetylcholinesterase and carbonic anhydrase activity. Drug and Chemical Toxicolology, 42(6):634-640.
• Balaydın HT, Özil M, Şentürk M, 2018. Synthesis and glutathione reductase inhibitory properties of 5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one's aryl Schiff base derivatives. Archiv der Pharmazie, e1800086.
• Beydemir S, Demir Y, 2017. Antiepileptic drugs: impacts on human serum paraoxonase-1. Journal of Biochemical and Molecular Toxicology, 31(6): e21889.
• Bradford MM, 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248–254.
• Budak H, Kocpinar EF, Gonul N, Ceylan H, Erol HS, Erdogan O, 2014. Stimulation of gene expression and activity of antioxidant-related enzyme in Sprague Dawley rat kidney induced by long-term iron toxicity. Comparative Biochemistry and Physiology - Part C, 166: 44–50.
• Caglayan C, Demir Y, Kücükler S, Taslimi P, Kandemir FM, Gulcin, I, 2018. The Effects of Hesperidin on Sodium Arsenite-Induced Different Organ Toxicity in Rats on Metabolic Enzymes as Antidiabetic and Anticholinergics Potentials: A Biochemical Approach. Journal of Food Biochemistry, 43(2): e12720.
• Carlberg I, Mannervik B, 1985. Methods Enzymol Academic Press, Orlando FL.
• Ceylan H, Demir Y, Beydemir, Ş, 2019. Inhibitory effects of usnic and carnosic acid on some metabolic enzymes: an in vitro study. Protein Peptide Letters, 26(5):364-370.
• Demir Y, Beydemir S, 2015. Purification, refolding, and characterization of recombinant human paraoxonase-1. Turkish Journal of Chemistry, 39: 764–776.
• Demir Y, Dikbaş N, Beydemir Ş, 2018b. Purification and Biochemical Characterization of Phytase Enzyme from Lactobacillus coryniformis (MH121153) Molecular Biotechnology, 60(11): 783-790.
• Demir Y, Isık M, Gulcin I, Beydemir S, 2017a. Phenolic compounds inhibit the aldose reductase enzyme from the sheep kidney. Journal of Biochemical and Molecular Toxicology, 31(9): e21935.
• Demir Y, Kotan M, Dikbaş N, Beydemir Ş, 2017b. Phytase from Weissella halotolerans: Purification, partial characterization and the effect of some metals. International Journal of Food Properties, 20(2): 2127-2137.
• Demir Y, Köksal, Z, 2019. The inhibition Effects of Some Sulfonamides on Human Serum Paraoxonase-1 (hPON1), Pharmacological Reports, 71(3):545-549.
• Demir Y, Oruç E, Topal A, 2016. Carbonic Anhydrase Activity Responses and Histopathological Changes in Gill and Liver Tissues after Acute Exposure to Chromium in Brown Trout Juveniles. Hacettepe Journal of Biology and Chemistry, 44 (4): 515–523.
• Demir Y, Taslimi P, Ozaslan MS, Oztaskin N, Çetinkaya Y, Gulçin İ, Beydemir Ş, Goksu S, 2018a. Antidiabetic potential: In vitro inhibition effects of bromophenol and diarylmethanones derivatives on metabolic enzymes. Archiv der Pharmazie, 351(12):e1800263.
• Erat M, Sakiroglu H, Ciftci M, 2005. Effects of some antibiotics on glutathione reductase activities from human erythrocytes in vitro and from rat erythrocytes in vivo. Journal of Enzyme Inhibition and Medicinal Chemistry, 20: 69-74.
• Farber NB, Jiang XP, Heinkel C, Nemmers B, 2002. Antiepileptic drugs and agents that inhibit voltage-gated sodium channels prevent NMDA antagonist neurotoxicity. Molecular Psychiatry, 7(7): 726-33.
• Heli H, Faramarzi F, Sattarahmady N, 2012. Oxidation and determination of Gabapentin on nanotubes of nickel oxide-modified carbon paste electrode, Journal of Solid State Electrochemistry, 16(1): 45-52.
• Işık M, Demir Y, Kırıcı M, Demir R, Şimşek F, Beydemir Ş, 2015. Changes in the anti-oxidant system in adult epilepsy patients receiving anti-epileptic drugs. Archives of Physiology and Biochemistry, 121: 97–102.
• Kırıcı M, Kırıcı M, Demir Y, Beydemir Ş, Atamanalp M, 2016. The Effect of Al3+ and Hg2+ on Glucose 6-Phosphate Dehydrogenase from Capoetaumbla kidney. Applied Ecology and Environmental Research, 14(2): 253-264.
• Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680–685.
• Lineweaver H, Burk D, 1934. The determination of enzyme dissociation constants. Journal of the American Chemical Society, 56: 658–666.
• Ozaslan MS, Demir Y, Aslan HE, Beydemir S, Kufrevioglu OI, 2018. Evaluation of chalcones as inhibitors of glutathione S-transferase. Journal of Biochemical and Molecular Toxicology, 32(5), e22047.
• Ozaslan MS, Demir Y, Kufrevioglu OI, Ciftci M, 2017. Some metals inhibit the Glutathione S-transferase from Van Lake fish gills. Journal of Biochemical and Molecular Toxicology, 31(11): e21967.
• Senturk M, Kufrevioglu OI, Ciftci M, 2008. Effects of some antibiotics on human erythrocyte glutathione reductase: an in vitro study. Journal of Enzyme Inhibition and Medicinal Chemistry, 23(1): 144–148.
• Taslimi P, Aslan HE, Demir Y, Oztaskin N, Maraş A, Gulçin İ, Beydemir S, Goksu S, 2018b. Diarylmethanon, bromophenol and diarylmethane compounds: Discovery of potent aldose reductase, α-amylase and α-glycosidase inhibitors as new therapeutic approach in diabetes and functional hyperglycemia. International Journal of Biological Macromolecules, 119: 857-863.
• Taslimi P, Kandemir FM, Demir Y, Ileritürk M, Temel Y, Caglayan C, Gulcin, I, 2018a. The Antidiabetic and Anticholinergic Effects of Chrysin on Cyclophosphamide-Induced Multiple Organs Toxicity in Rats: Pharmacological Evaluation of Some Metabolic Enzymes Activities. Journal of Biochemical and Molecular Toxicology, 33(6):e22313.
• Türkan F, Huyut Z, Demir Y, Ertaş F, Beydemir Ş, 2018. The effects of some cephalosporins on acetylcholinesterase and glutathione S-transferase: an in vivo and in vitro study. Archives of Physiology and Biochemistry, 125(3):235-243.
• Türkeş C, Demir Y, Beydemir S, 2019. Anti-diabetic Properties of Calcium Channel Blockers: Inhibition Effects on Aldose Reductase Enzyme activity. Applied Biochemistry and Biotechnology, 189(1):318-329.
• Yamatogi Y, 2004. Principles of antiepileptic drug treatment of epilepsy. Psychiatry and Clinical Neurosciences, 58(3): 3-6.