SCHISTOSOMA-MANSONI - SINGLE-STEP PURIFICATION AND CHARACTERIZATION OF GLUTATHIONE-S-TRANSFERASE ISOENZYME-4

被引:26
作者
OLEARY, KA
HATHAWAY, KM
TRACY, JW
机构
[1] UNIV WISCONSIN,DEPT COMPARAT BIOSCI,2015 LINDEN DR W,MADISON,WI 53706
[2] UNIV WISCONSIN,DEPT PHARMACOL,MADISON,WI 53706
[3] UNIV WISCONSIN,CTR ENVIRONM TOXICOL,MADISON,WI 53706
关键词
TREMATODE; SCHISTOSOMA-MANSONI; ISOENZYME; AFFINITY CHROMATOGRAPHY; SUBSTRATE SPECIFICITY; 4-HYDROXYALK-2-ENAL; CUMENE HYDROPEROXIDE; H2O2;
D O I
10.1016/0014-4894(92)90121-P
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
A soluble glutathione S-transferase isoenzyme, designated SmGST-4 was purified to apparent homogeneity in a single step from the cytosol of adult Schistosoma mansoni by selective elution of the enzyme from a glutathione-agarose affinity column using glutathione disulfide. SmGST-4, which comprised about 5% of the bound glutathione S-transferase activity, could be distinguished from the previously characterized glutathione S-transferase isoenzyme family (SmGST-1/2/3) by its unique Chromatographic behavior, lower subunit Mr (26,000), differences in substrate specificity and inhibitor sensitivity, and a lack of reactivity with antiserum to SmGST-3. The purified isoenzyme catalyzed the conjugation of several model xenobiotics including 1-chloro-2,4-dinitrobenzene, ethacrynic acid, and trans-4-phenyl-3-buten-2-one. Like the SmGST-1/2/3 isoenzyme family, SmGST-4 failed to catalyze the conjugation of a model epoxide substrate, 1,2-epoxy-3-(p-nitrophenoxy)propane. Because glutathione S-transferases from other organisms play a role in protecting cells against the toxic products of lipid peroxidation, SmGST-4 and the members of the SmGST-1/2/3 isoenzyme family were tested for their capacity to reduce cumene hydroperoxide and to catalyze the conjugation of 4-hydroxyalk-2-enals. Although all four isoenzymes catalyzed both reactions, the specific activity of SmGST-1, SmGST-2, and SmGST-3 toward cumene hydroperoxide was at least 10-fold greater than that of SmGST-4. In contrast, the latter more effectively conjugated a homologous series of 4-hydroxyalk-2-enal isomers. For example, the specific activity of SmGST-4 toward 4-hydroxynon-2-enal (10.8 μmol · min-1 · mg protein-1), the isomer formed under physiological conditions, was about 5-fold greater than that of SmGST-1 and 12-fold greater than that of the major isoenzyme SmGST-3. These results are consistent with a multifunctional role for schistosome glutathione 5-transferases in protecting the parasite against both xenobiotics and toxic endogenous products of lipid peroxidation. © 1992.
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页码:47 / 55
页数:9
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