title: Chemical modification of human placental glutathione transferase by pyridoxal 5'-phosphate.
abstract: Incubation of GST pi from human placenta with 8 mM PLP resulted in a rapid loss of activity during the first 10 min, concomitant with a Schiff base formation. This inactivation was probably due to the formation of a reversible adduct between PLP and the enzyme. After sodium borohydride treatment this adduct was reduced and stabilized. Stoichiometry and peptide isolation studies showed that three lysine residues were modified during reaction of GST and PLP. Protection of the enzyme against inactivation was achieved in the presence of 4 mM GSH suggesting that at least one lysyl residue is associated with the substrate binding site. Peptide mapping by digesting the enzyme with trypsin revealed that lysine shielded by GSH is Lys-127. Our results suggest that this residue may play an important role in enzymatic activity.
authors.affiliation: Department of Biology, 2nd University of Rome Tor Vergata, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Lo Bello
authors.forename: R
authors.initials: R
authors.lastname: Petruzzelli
authors.forename: L
authors.initials: L
authors.lastname: Reale
authors.forename: G
authors.initials: G
authors.lastname: Ricci
authors.forename: D
authors.initials: D
authors.lastname: Barra
authors.forename: G
authors.initials: G
authors.lastname: Federici
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D011732:Pyridoxal Phosphate / Q000378:metabolism / Q000494:pharmacology*; D013053:Spectrophotometry; D014357:Trypsin
chemical_list: D007527:Isoenzymes; D010446:Peptide Fragments; D011732:Pyridoxal Phosphate; D005982:Glutathione Transferase; D014357:Trypsin