title: Purification and characterization of five forms of glutathione transferase from human uterus.
abstract: Five glutathione transferase (GST) forms were purified from human uterus by glutathione-affinity chromatography followed by chromatofocusing, and their structural, kinetic and immunological properties were investigated. Upon SDS/polyacrylamide slab gel electrophoresis all forms resulted composed of two subunits of identical molecular size. GST V (pI 4.5) is a dimer of 23-kDa subunits. GST I (pI 6.8) and GST IV (pI 4.9) are dimers of 24-kDa subunits whereas GST II (pI 6.1) and GST III (pI 5.5) are dimers of 26.5-kDa subunits. GST V accounts for about 85-90% of the activity whereas the other isoenzymes are present in trace quantities. On the basis of the molecular mass of the subunits, amino acid composition, substrate specificities, sensitivities to inhibitors, CD spectra and immunological studies, GST V appeared very similar to transferase pi. Structural and immunological studies provide evidence that GST IV is closely related to the less 'basic' transferase (GST pI 8.5) of human skin. Extensive similarities have been found between GST II and GST III. The comparison includes amino acid compositions, subunits molecular size and immunological properties. The two enzymes, however, are kinetically distinguishable. The data presented also indicate that GST II and GST III are related to transferase mu and to transferase psi of human liver. Even though GST I has a subunit molecular mass identical to GST IV, several lines of evidence, including catalytic and immunological properties, indicate that they are different from each other. GST I seems not to be related to any of known human transferases, suggesting that it may be specific for the uterus.
authors.affiliation: Istituto di Scienze Biochimiche, Facoltà di Medicina, Università G. D'Annunzio, Chieti, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Di Ilio
authors.forename: A
authors.initials: A
authors.lastname: Aceto
authors.forename: G
authors.initials: G
authors.lastname: Del Boccio
authors.forename: E
authors.initials: E
authors.lastname: Casalone
authors.forename: A
authors.initials: A
authors.lastname: Pennelli
authors.forename: G
authors.initials: G
authors.lastname: Federici
mesh_terms: D000345:Affinity Labels; D000596:Amino Acids / Q000302:isolation & purification; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005978:Glutathione; D005982:Glutathione Transferase / Q000302:isolation & purification*; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D014599:Uterus / Q000201:enzymology*
chemical_list: D000345:Affinity Labels; D000596:Amino Acids; D005982:Glutathione Transferase; D005978:Glutathione