Molecular cloning, expression and characterization of a recombinant glutathione S-transferase from Echinococcus multilocularis

被引：22

作者：

Liebau, E ^{[1
]}

Muller, V ^{[1
]}

Lucius, R ^{[1
]}

Walter, RD ^{[1
]}

HenkleDuhrsen, K ^{[1
]}

机构：

[1] UNIV HOHENHEIM, DEPT ZOOL, DIV PARASITOL, D-70593 STUTTGART, GERMANY

来源：

MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 1996年 / 77卷 / 01期

关键词：

glutathione S-transferase; cestode; Echinococcus multilocularis;

D O I：

10.1016/0166-6851(96)02578-9

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the identification and characterization of the first cestode glutathione S-transferase (GST) cDNA sequence. A fragment of an Echinococcus multilocularis glutathione S-transferase cDNA was isolated by the polymerase chain reaction. Subsequently, a Lambda zap cDNA library prepared from mRNA from protoscolices of E. multilocularis was screened with this PCR fragment. A complete cDNA clone was isolated and the nucleotide sequence determined. Analysis of the E. multilocularis GST-deduced amino acid sequence indicates that it is clearly related to the mammalian mu-class GSTs. The E. multilocularis GST cDNA was expressed in Escherichia coli, using a protocol designed to produce the native enzyme rather than a fusion protein. The 25.5-kDa enzyme subunit was purified to homogeneity using glutathione-sepharose chromatography. Gel filtration demonstrated that this GST is enzymatically active as a homodimer. The recombinant enzyme had conjugating activity with organic hydroperoxides and with members of the trans,trans-2,4 alkadienal and trans-2-alkenal series, which are secondary products of lipid peroxidation.

引用

页码：49 / 56

页数：8

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