title: Purification of Pumpkin Glutathione S-Transferase Species Specifically Present in Cultured Cells Treated by Excessive Concentration of 2,4-Dichlorophenoxyacetic Acid but Absent in Normal Plants.
abstract: Pugc is a unique glutathione S-transferase subunit species that is absent in normal pumpkin plants, but GST3c (homodimer of Pugc) accumulates in cultured pumpkin cells treated with 40 ppm 2,4-dichlorophenoxyacetic acid for six days. GST3c was purified by DEAE-cellulose, hydroxylapatite, and S-hexylglutathione-agarose column chromatography, and its homogeneity was confirmed by SDS-PAGE analysis. The specific activity of GST3c was 124 μmol/min/mg protein with 1-chloro-2,4-dinitrobenzene. This was higher than those of the other plant and animal glutathione S-transferases. Mouse antiserum raised against the purified GST3c recognized only Pugc, but not Puga, Pugb, nor Pugd when the reactivity of pumpkin glutathione S-transferase subunits was tested.
authors.affiliation: a Department of Plant Sciences, Faculty of Agriculture, Kagawa University.
authors.forename: M
authors.initials: M
authors.lastname: Fujita
authors.affiliation: a Department of Plant Sciences, Faculty of Agriculture, Kagawa University.
authors.forename: Y
authors.initials: Y
authors.lastname: Adachi
authors.affiliation: a Department of Plant Sciences, Faculty of Agriculture, Kagawa University.
authors.forename: N
authors.initials: N
authors.lastname: Sakato
keywords: 2,4-dichlorophenoxyacetic acid; antiserum; glutathione S-transferase; pumpkin; purification