Analytical Science and Technology (분석과학)

The Korean Society of Analytical Sciences (한국분석과학회)
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Analysis on the substrate specificity and inhibition effect of Brassica oleracea glutathione S-Transferase

양배추 유래의 글루타티온 전달효소의 기질 특이성 및 저해 효과 분석

  • Park, Hee-Joong (Department of Chemistry, College of Natural Sciences, Chung-Ang University) ;
  • Lee, Hee-Jin (Department of Chemistry, College of Natural Sciences, Chung-Ang University) ;
  • Kong, Kwang-Hoon (Department of Chemistry, College of Natural Sciences, Chung-Ang University)
  • 박희중 (중앙대학교 자연과학대학 화학과) ;
  • 이희진 (중앙대학교 자연과학대학 화학과) ;
  • 공광훈 (중앙대학교 자연과학대학 화학과)
  • Received : 2009.03.05
  • Accepted : 2009.06.01
  • Published : 2009.06.25
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Abstract

To gain further insight into herbicide detoxification of plant, we purified a glutathione S-transferase from Brassica oleracea (BoGST) and studied its substrate specificity towards several xenobiotic compounds. The BoGST was purified to electrophoretic homogeneity with approximately 10% activity yield by DEAE-Sephacel and GSHSepharose column chromatography. The molecular weight of the BoGST was determined to be approximately 23,000 by SDS-polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the BoGST was significantly inhibited by S-hexyl-GSH and S-(2,4-dinitrophenyl)GSH. The substrate specificity of the BoGST displayed high activities towards CDNB, a general GST substrate and ethacrynic acid. It also exhibited GSH peroxidase activity toward cumene hydroperoxide.

본 연구에서는 식물의 제초제 해독 기구를 알아보기 위해, 양배추 (Brassica oleracea)로 부터 글루타티온 전달효소를 정제하고 외래성 이물질에 대한 기질 특이성과 저해 효과를 분석하였다. 양배추로부터 DEAE-Sephacel 컬럼과 GSH-Sepharose 컬럼 크로마토그래피를 이용하여 약 10%의 수율로 글루타티온 전달효소를 정제하였다. 정제된 효소에 대해 분자량을 측정한 결과, SDS-polyacrylamide 겔 전기영동으로 측정한 분자량은 23,000Da을 나타내었으며, 겔 크로마토그래피로 측정한 분자량은 48,000Da을 나타내었다. 따라서 정제된 양배추 글루타티온 전달효소는 소단위체의 분자량이 약 23,000Da의 동종이량체라는 사실을 알 수 있었다. 이 효소의 저해제에 대한 효과를 조사한 결과, S-hexyl-GSH와 S-(2,4-dinitrophenyl)GSH에 의해 활성이 저해되었다. 양배추 글루타티온 전달효소의 기질특이성은 CDNB와 ETA에서 높은 활성을 보였으며, cumene hydroperoxide에 대한 GSH peroxidase 활성도 나타내었다.

Keywords

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