title: Non-essentiality of cysteine and histidine residues for the activity of human class PI glutathione S-transferase.
abstract: In order to examine the roles of cysteine and histidine residues in the activity of human class Pi glutathione S-transferase (GST pi), site-directed mutagenesis was used to replace each of the four cysteine residues (at positions 14, 47, 101 and 169) with serine and each of the two histidine residues (at positions 71 and 162) with asparagine using a cDNA for the enzyme (Kano, T. et al. (1987) Cancer Res., 47, 5626-5630) and an E. coli expression system. The replacements of Cys101, Cys169, His71 and His162 did not affect the GSH-conjugating activity toward 1-chloro-2,4-dinitrobenzene and ethacrynic acid. On the other hand, the activities were partly decreased by the replacements of Cys47 and Cys14. These results indicated that the cysteine and histidine residues in GST pi are not essential for the catalytic activity, although Cys47 and Cys14 may contribute to some extent to the catalytic efficiency.
authors.affiliation: Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
authors.forename: K H
authors.initials: KH
authors.lastname: Kong
authors.forename: H
authors.initials: H
authors.lastname: Inoue
authors.forename: K
authors.initials: K
authors.lastname: Takahashi
mesh_terms: D001483:Base Sequence; D003545:Cysteine / Q000235:genetics; D004247:DNA / Q000235:genetics; D004137:Dinitrochlorobenzene / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D004976:Ethacrynic Acid / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D004137:Dinitrochlorobenzene; D006639:Histidine; D004247:DNA; D005982:Glutathione Transferase; D003545:Cysteine; D004976:Ethacrynic Acid