title: Purification and characterization of a novel alpha-class glutathione transferase from human liver.
abstract: The importance of glutathione transferases (GST) as a major group of detoxification enzymes is well known. The human liver possesses these enzymes in high concentration and in a multiplicity of forms. We describe here a novel glutathione transferase isoenzyme isolated from liver using glutathione affinity chromatography, DEAE-sepharose and Mono-Q ion-exchange chromatography. The isoenzyme is a dimer of approximately 25 kDa with a blocked N-termini. Its kinetic and immunological properties indicate that it belongs to the alpha-class of GSTs. Its isoelectric point (8.0) is closely related to GST alpha (pI 7.8) and GST beta (pI 8.2) reported previously. More than 70% of the amino-acid sequence of this isoenzyme has been determined by automated Edman degradation procedure. The results suggest that this isoenzyme (which we term GST 8.0) may be a heterodimer of two, closely related, novel alpha-class GST subunits. Comparisons between the amino acid sequences of these two novel alpha-class subunits with those of the other alpha-class GST subunits already known indicate changes in a number of different residues localized in the electrophilic binding site. Further studies are needed to establish whether such differences are due to allelic polymorphism of the enzyme or to the existence of additional genes for alpha-class GSTs in human liver. These results are consistent with previous data which suggest that a multitude of different GSTs, especially of alpha class, are present in the human liver providing this tissue with an efficient mechanism of protection against xenobiotic and endogenous compounds.
authors.affiliation: Laboratory of Clinical Biochemistry, Bambino Gesù Children's Hospital, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Pastore
authors.forename: M
authors.initials: M
authors.lastname: Lo Bello
authors.forename: G
authors.initials: G
authors.lastname: Aureli
authors.forename: G
authors.initials: G
authors.lastname: Federici
authors.forename: G
authors.initials: G
authors.lastname: Ricci
authors.forename: C
authors.initials: C
authors.lastname: Di Ilio
authors.forename: R
authors.initials: R
authors.lastname: Petruzzelli
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D015153:Blotting, Western; D003429:Cross Reactions; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes; D008099:Liver / Q000737:chemistry / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D013379:Substrate Specificity
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase