Purification and characterization of a Cu, Zn-superoxide dismutase from adult <i>Paragonimus westermani</i>

, Chung, Y B; , Song, C Y; , Lee, H S; , Kong, Y; , Cho, S Y

doi:1991.29.3.259

Korean J Parasito > Volume 29(3):1991 > Article

Original Article


Korean J Parasitol. 1991 Sep;29(3):259-266. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1991.29.3.259
Copyright © 1991 by The Korean Society for Parasitology


Purification and characterization of a Cu, Zn-superoxide dismutase from adult Paragonimus westermani
Y B Chung,C Y Song,H S Lee,^Y Kong,^* and S Y Cho^**
Department of Biology, College of Natural Sciences, Chung-Ang University, Seoul 156-756, Korea.


Abstract
In cytosolic fraction of adult Paragonimus westermani, superoxide dismutase activity was identified (4.3 units/mg of specific activity) using a xanthine-xanthine oxidase system. The enzyme was purified 150 fold in its activity using the ammonium sulfate precipitation, DEAE-Trisacryl M anion-exchange chromatography and Sephadex G-100 molecular sieve chromatography. The enzyme exhibited the enhanced activity at pH 10.0. The enzyme activity totally disappeared in 1.0mM cyanide while it remained 77.8% even in 10 mM azide. These findings indicated that the enzyme was Cu, Zn-SOD type. Molecular mass of the enzyme was estimated to be 34 kDa by gel filtration and 17 kDa on reducing SDS-polyacrylamide gel electrophoresis which indicated a dimer protein.

Figures


	Fig. 1 Elution profile of SOD on the 1st DEAE-Trisacryl1 M anion-exchange chromatography. An allocation of each fraction was assayed form SOD activity (○) and protein (•). A linear gradient was indicated by dotted line. Pooled fractions containing SOD activity was shown by a bar (-).


	Fig. 2 Elution profile of SOD on the 2nd DEAE-Trisacry1 M anion-exchange chromatography. Markings are the same as described in Fig. 1.


	Fig. 3 Elution profile of SOD on the 3nd DEAE-Trisacry1 M anion-exchange chromatography. Markings are the same as described in Fig. 1.


	Fig. 4 Eatimation of molecular mass of Cu, Zn-SOD by Sephadex G-100 gel filtration. A : Bovine serum albumin (68 kDa) B : Ovalbumin (43 kDa) C : Chymotrypsinogen A (25 kDa) D : Cytochrome c (12.4 kDa)


	Fig. 5 Findings on SDS-PAGE of Cu, Zn-SOD of P. westermani. The samples were electrophoresed at 12.5% gel in reducing condition. Mr : Molecular mass in kDa C : Crude extract 1 : 1st DEAE-Trisacryl M anion-exchange chromatography 2 : 2nd DEAE-Trisacryl M anion-exchange chromatography 3 : 3rd DEAE-Trisacryl M anion-exchange chromatography 4 : Sephadex G-100 column chromatography

Tables


	Table 1 Purification of cytosolic superoxide dismutase from P. westermani


	Table 2 Effects of inhibitors on activity of cytosolic SOD of P. westermani


	Table 3 Effects of pH on activity of cytosolic SOD of P. westermani

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