Biochemical properties of cloned glutathione S-transferases from Schistosoma mansoni and Schistosoma japonicum.
cDNA clones encoding a 28-kDa subunit glutathione S-transferase (GST) from Schistosoma mansoni (Sm28GST) and a 26-kDa subunit GST from Schistosoma japonicum (Sj26GST) have been expressed in bacterial systems. The recombinant proteins were purified to homogeneity by batch-wash glutathione-agarose affinity chromatography and their biochemical properties investigated. Gel filtration chromatography indicated that both recombinant GSTs are homodimeric proteins. Resolution of Sm28GST and Sj26GST by chromatofocusing in the ranges pH 9-6 and pH 7-4 gave pI estimates of 7.4 and 5.0, respectively. Kinetic analyses suggested that both Sm28GST and Sj26GST operate via a sequential bisubstrate catalytic mechanism. Sm28GST and Sj26GST displayed a mosaic of mammalian Alpha-, Mu- and Pi-type substrate specificities and inhibitor sensitivities. However, multivariate analysis suggests that Sm28GST has an overall catalytic homology with mammalian Mu class GSTs, whilst the enzymatic properties of Sj26GST appear to constitute a hybridisation of Mu and Alpha class features. Both recombinant GSTs interact with a range of hydrophobic ligands including haematin and related compounds, bile acids and several anthelmintics. Sm28GST and Sj26GST possess relatively limited selenium-independent glutathione peroxidase activities, but are able to catalyse the glutathione conjugation of members of the trans,trans-alka-2,4-dienal, trans-alk-2-enal and 4-hydroxyalk-2-enal series of reactive carbonyls (known secondary products of lipid peroxidation).
Walker
J
J
Department of Biological Sciences, University of Wales at Aberystwyth, Dyfed, UK.
Crowley
P
P
Moreman
A D
AD
Barrett
J
J
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D016054:DNA, Protozoan / Q000378:metabolism; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology*; D012550:Schistosoma mansoni / Q000201:enzymology*; D013379:Substrate Specificity
D016054:DNA, Protozoan; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D005982:Glutathione Transferase