Purification of glucose 6-phosphate dehydrogenase from Buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties

Çiftçi, Mehmet; Beydemır, Şükrü; Yilmaz, H. Birkan; Altıkat, Sayit

doi:10.1016/s1046-5928(03)00073-1

Cited by 32 publications

(14 citation statements)

References 16 publications

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“…According to these results, GR is the most resistant enzyme against temperature. This result is also in parallel with similar results in the literature 16,35,[45][46][47][48] . For calculation of stable pH of enzymes, 200 mM phosphate buffers with 5.5, 6.0, 6.5, 7.0 and 7.5 pH, and 200 mM Tris-HCl buffers with 8.0, 8.5 and 9.0 pH were preserved at +4 C together with the equal amount of enzyme solution, and activity measurements were performed for 72 h with 12 h intervals.…”

Section: Discussionsupporting

confidence: 92%

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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: 92%

“…These specific activities, % efficiency and purification coefficients are optimal values for kinetic studies. Moreover, compared to the literature, the purification process was fairly successful and suitable for scientific studies 13,[35][36][37] . These enzymes have been purified from several mammalian tissues.…”

Section: Discussionmentioning

confidence: 99%

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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“…According to these values, the K M value for G6P is lower than that for NADP + indicating thus the affinity of G6P to the enzyme is greater than that of NADP + . Similar values were reported for G6PD from other sources [20][21][22] . Moreover, Cd +2 , Pb…”

Section: Resultssupporting

confidence: 88%

“…The optimum pH was found to be 8.0 in 50 mM Tris-HCl buffer. The result is similar to those of G6PDs obtained from different organisms [20][21][22] . Enzyme activities were measured between 0 and 90°C in order to search how temperature affected the enzyme.…”

Section: Resultssupporting

confidence: 87%

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In vitroenzymatic response of Turkish native chicken “Gerze” to heavy metal exposure

Mercan

Okumuş

Şentürk

et al. 2011

Journal of Enzyme Inhibition and Medicinal Chemistry

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Antibiotic effects on SO uptake in human erythrocytes

Casella

Pace

Romano

et al. 2007

Cell Biochemistry & Function

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The erythrocyte is a cell highly exposed to oxygen pressure that, in turn, provokes oxidative stress involving loss of SH-groups, cell shrinkage by activation of K(+)-Cl(-) cotransport (KCC) and membrane destabilization which plays an important role in the premature haemolysis of red blood cells (RBCs). Oxidative stress provoked by chemicals frequently occurs in human erythrocytes. The aim of this study was to test whether the antibiotics alter the redox state and investigate their influences on band 3 protein that is involved in the facilitated electro neutral exchange of Cl(-) for HCO(3)(-) across the membrane of mammalian erythrocytes. Normal erythrocytes were treated with some antibiotics and thiol oxidizing agent N-ethylmaleimide (NEM) and tested for sulphate uptake, K(+) efflux and for glutathione (GSH) concentration as an index of oxidative stress. The rate constant of SO(4)(=) uptake measured in erythrocytes treated with antibiotics as well as NEM was decreased with respect to control cells as a result of band 3 SH-groups oxidation or the stress-induced K(+)-Cl(-) symport-mediated cell shrinkage. In fact, this hypothesis was verified by increased K(+) efflux and decreased GSH values measured in treated erythrocytes compared to controls.

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Purification of glucose 6-phosphate dehydrogenase from Buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties

Cited by 32 publications

References 16 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

In vitroenzymatic response of Turkish native chicken “Gerze” to heavy metal exposure

Antibiotic effects on SO uptake in human erythrocytes

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