Sheep brain glutathione reductase: Purification and general properties

Açan, N. Leyla; Tezcan, E. Ferhan

doi:10.1016/0014-5793(89)80687-8

Cited by 49 publications

(17 citation statements)

References 10 publications

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“…Hence, these pH values are accepted as optimal pH values. Research showed that these results are highly parallel to other results in the literature 8,37,[42][43][44] . For optimal temperature calculation of the enzymes, activities of optimal ionic strength and optimal pH enzymes were measured at 10 C intervals within 10-70 C range.…”

Section: Discussionsupporting

confidence: 82%

Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics

Adem

Çiftçi

2016

Journal of Enzyme Inhibition and Medicinal Chemistry

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G6PD, 6PGD and GR have been purified separately in the single step from rat lung using 2 0 , 5 0 -ADP Sepharose 4B affinity chromatography. The purified enzymes showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of the enzymes were estimated to be 134 kDa for G6PD, 107 kDa for 6PGD and 121 kDa for GR by Sephadex G-150 gel filtration chromatography, and the subunit molecular weights was respectively found to be 66, 52 and 63 kDa by SDS-PAGE. Optimum pH, stable pH, optimum ionic strength, optimum temperature, K M and V max values for substrates were determined. Product inhibition studies were also performed. The enzymes were inhibited by levofloxacin, furosemide, ceftazidime, cefuroxime and gentamicin as in vitro with IC 50 values in the range of 0.07-30.13 mM. In vivo studies demonstrated that lung GR was inhibited by furosemide and lung 6PGD was inhibited by levofloxacin.

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Section: Discussionsupporting

confidence: 82%

Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics

Adem

Çiftçi

2016

Journal of Enzyme Inhibition and Medicinal Chemistry

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“…Little is known about the biochemical properties and the localization of GR in brain. So far the purification of GR has been reported for sheep brain (Acan and Tezcan, 1989). The kinetic parameters of the purified GR (Acan and Tezcan, 1991) and the inhibition of the enzyme by Cd 2+ (Acan and Tezcan, 1995) have been published.…”

mentioning

confidence: 99%

Purification of Glutathione Reductase from Bovine Brain, Generation of an Antiserum, and Immunocytochemical Localization of the Enzyme in Neural Cells

Gutterer

Dringen

Hirrlinger

et al. 1999

Journal of Neurochemistry

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Glutathione reductase (GR) is an essential enzyme for the glutathione-mediated detoxification of peroxides because it catalyzes the reduction of glutathione disulfide. GR was purified from bovine brain 5,000-fold with a specific activity of 145 U/mg of protein. The homogeneity of the enzyme was proven by sodium dodecyl sulfate -polyacrylamide gel electrophoresis and silver staining of the gel. The purified GR from bovine brain is a dimer of two subunits that have an apparent molecular mass of 55 kDa. The purified GR was used to generate a rabbit antiserum with the intention to localize GR in brain cells. The antiserum was useful for the detection of GR by double-labeling immunocytochemical staining in astroglia-rich and neuron-rich primary cultures from rat brain. In homogenates of these cultures, no significant difference in the specific activities of GR was determined. However, not all cell types present in these cultures showed identical staining intensity for GR. In astrogliarich primary cultures, strong GR immunoreactivity was found in cells positive for the cellular markers galactocerebroside and C3b (antibody Ox42), indicating that oligodendroglial and microglial cells, respectively, contain GR. In contrast, only weak immunoreactivity for GR was found in cells positive for glial fibrillary acidic protein.In neuron-rich primary cultures, GAP43-positive cells stained with the antiserum against GR. These data demonstrate that, in cultures of neural cells, neurons, oligodendroglial cells, and microglial cells express high levels of GR. Key Words: Antiserum-Astroglia-Brain-Glutathione-Microglia-Oligodendroglia-Neurons.

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“…The isoenzymes were eluted with a gradient of 0-0.5 mM GSH and 0-1 mM NADPH in 50 mM Kphosphate, containing 1 mM EDTA (pH 7.3). Active fractions were collected and dialyzed with equilibration buffer at 4 C (Acan & Tezcan, 1989;Boggaram et al, 1979;Carlberg & Mannervik, 1981).…”

Section: Preparation Of the Hemolysatementioning

confidence: 99%

The effect of some antibiotics on glutathione reductase enzyme purified from liver and erythrocyte of Lake Van pearl mullet

Altun

Türkoğlu²,

Çelik

2015

Pharmaceutical Biology

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Context: The effect of antibiotics (amikacin, cefazolin, ivermectin, and kanamycin) on glutathione reductase (GR) isoenzymes activity in liver and erythrocyte of the fish, Chalcalburnus tarichi (Lake Van pearl mullet, Pallas 1811) (Cyprinidae) were investigated. Objective: This study determined the biochemical characterization of GR purified from the liver and erythrocytes of C. tarichi and the inhibition effect of the antibiotics on the GR isoenzymes. Materials and methods: GR was purified by affinity chromatography from the tissues of C. tarichi. The biochemical characterization of GR such as optimum temperature, optimum pH, and ionic strength were determined. The inhibition effects of the antibiotics on the isoenzymes were evaluated as IC 50 and K i values. K i constant and 50% inhibitory concentration (IC 50 ) value for antibiotics were determined by Lineweaver-Burk graphs and plotting activity % versus [I], respectively, at five different concentrations of antibiotics. Results: Optimum temperature, pH, and ionic strength were determined for isoenzymes as 40 C, 60 C; 8.0, 8.0, and 50, 50 mM, respectively. Subunit molecular weights of the isoenzymes were estimated as 55 kDa by sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE). In addition, IC 50 and K i values were calculated for amikacin, cefazolin, ivermectin, and kanamycin. The antibiotics showed non-competitive inhibition effects. IC 50 values were calculated as 16.3, 36.6, 0.504, and 18.8 mM for liver and 20.0, 30.4, 0.787, and 31

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Sheep brain glutathione reductase: Purification and general properties

Cited by 49 publications

References 10 publications

Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics

Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics

Purification of Glutathione Reductase from Bovine Brain, Generation of an Antiserum, and Immunocytochemical Localization of the Enzyme in Neural Cells

The effect of some antibiotics on glutathione reductase enzyme purified from liver and erythrocyte of Lake Van pearl mullet

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