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Purification and Characterization of Glutathione S-transferase π from Human Placental Tissues
인체 태반조직 Glutathione S-transferase π의 정제 및 특성구명

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Authors
Kwak, Sahng-June; Park, Sang Chul
Issue Date
1988-06
Publisher
Seoul National University College of Medicine
Citation
Seoul J Med, Vol.29 No.2, pp. 107-118
Keywords
Human placental glutathione S-transferase(GST- π)PurificationCharacterization
Abstract
Human placental glutathione S-transferase(GST - π ) was purified to the apparent
homgeneity through salting-out with ammonium sulfate and the consecutive chromatography
on carboxymethyI(CM)-, diethylaminoethyI(DEAE)-cellulose and S-hexylglutathione
sepharose 6B affinity column. For the characterization of the apparently purified enzyme,
sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS PAGE), kinetic studies, neuraminidase
digestion, and isoelectrofocusing were performed. The yield of the enzyme was
11 percent with the 1107 fold purification and the respective specific activity to
l-chloro-2,4-dinitrobenzene(CDNB), 1,2-dichloro-4-nitrobenzene(DCNB) and p-nitrophenyl
chloride was 62 JU/mg, 0.12 IU/mg, and almost non-detectable. The Km of the enzyme for reduced
glutathione(GSH) was 0.085 mM at the concentration of 2 mM CDNB, while its Km for
CDNB was 0.46 mM at the fixed concentration of 5 mM GSH. Calcium, magnesium, zinc,
ethylenediamintertraacetic acid(EDTA) and ethylenedioxydiethylenedinitrilotetraacetic acid
(EGTA) did not show any significant effect on enzyme activity. The subunit of the enzyme
with a molecular weight of 25,000 did not reveal the molecular weight change after neuraminidase
treatment. Isoelectrofocusing of the enzyme showed two bands, of which the pi of the
major band was 4.48, while that of the minor band, 4.55. The specific antibody, raised against
the purified GST- π in the rabbit serum indicated the immunologic cross-reactivity to the acidic
GST from the human granulocyte.
ISSN
0582-6802
Language
English
URI
http://hdl.handle.net/10371/6364
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College of Medicine/School of Medicine (의과대학/대학원)Dept. of Medicine (의학과)The Seoul Journal of MedicineThe Seoul Journal of Medicine Vol. 29 No.2 (1988)
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