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.
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.
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 yie...
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.
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.
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제안 방법
In this study, a GST from the leaves of cabbage (Brassica oleracea) was purified, characterized the biochemical properties and compared its properties with those of enzymes from other sources.
The inhibition parameters (I50) of various inhibitors, 5-hexyl-GSH, S-methyl-GSH, benastatin A, EIA and S-(2, 4-dinitrophenyl)GSH for the GSH-CDNB conjugating activity were determined under the standard assay conditions (Table 3). The I50 value of 5-hexyl-GSH and S-methyLGSH, a derivative of GSH, for the enzyme was approximately 14 M.
대상 데이터
5 kDa) and lysozyme (144 kDa). Coomassie Blue R-250 was used for staining.
5% gel. Coomassie blue R-250 was used for staining. Lane M, molecular mass marker (Bio-Rad); lane 1, GSH-Sepharose column chromatography.
The leaves of cabbage (Brassica oleracea) used in this study were purchased from Huksuk market (produced in Seosan, Chungcheongnam-do). Reduced glutathione (GSH), dithiothreitol (DTT), ethylenediaminetetraacetic acid (EDTA), 1 -chloro-2, 4-dinitro- benzene (CDNB), le-dichloro-4-nitrobenzene (DCNB), ethacrynic acid (ETA), 1, 2-epoxy-3-(p-nitrophenoxy) propane (EPNP), DEAE-Sephacel and GSH-Sepharose were obtained from Sigma (St.
참고문헌 (36)
B. Mannervik and U. H. Danielson, CRC Crit. Rev. Biochem., 23, 283-337(1988)
B. Mannervik, Y. C. Awasthi, P. G. Board, J. D. Hayes, C. Ilio, B. Ketterer, I. Listowsky, R. Morgenstern, M. Muramatsu, W. R. Pearson, C. B. Pickett, K. Sato, M. Widersten and C. R. Wolf, Biochem. J., 282, 305-308(1992)
M. F. Lopez, W. F. Patton, W. B. Sawlivich, H. Erdjument-Bromage, P. Barry, K. Gmyrek, T. Hines, P. Tempst and W. M. Skea, Biochem. Biophys. Acta., 1205, 29-38(1994)
C. Di Ilio, A. Aceto, N. Allocati, R. Piccolomini, T. Bucciarelli, B. Dragani, A. Faraone, P. Sacchetta, R. Petruzzelli and G. Federici, Arch. Biochem. Biophys., 305, 110-114(1993)
C. Y. Lee, L. Johnson, R. H. Cox, J. D. McKinney and S. M. Lee, J. Biol. Chem., 256, 8110-8116(1981)
A. Aceto, B. Dragani, T. Bucciarelli, P. Sacchetta, F. Martini, S. Angelucci, F. Amicarelli, M. Miranda and C. Di Ilio, Biochem. J., 289, 417-422(1993)
K. H. Kong, H. Inoue and K. Takahashi, Biochem. Biophys. Res. Commun., 81, 748-755(1991)
P. Reinemer, L. Prade, P. Hof, T. Neuefeind, R. Huber, R. Zettl, K. Palme, J. Schell, I. Koelln, H. D. Bartunik and B. Bieseler, J. Mol Biol., 255, 289-309(1991)
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