The mechanism of the reaction between glutathione and 1-menaphthyl sulphate catalysed by a glutathione S-transferase from rat liver.
1. The glutathione S-transferase that catalyses the reaction of 1-menaphthyl (naphth-1-ylmethyl) sulphate with GSH was purified 76-fold from rat liver. 2. The properties of the purified enzyme were studied by gel filtration and isoelectric focusing. 3. The initial-velocity pattern in the absence of products and the product-inhibition pattern have been determined. These are consistent with an Ordered Bi Bi mechanism in which the GSH adds to the enzyme before 1-menaphthyl sulphate and the products are released in the order SO(4) (2-) followed by S-(1-menaphthyl)glutathione. 4. Dead-end-inhibition studies with p-aminobenzoic acid, which has been shown to be competitive with GSH and non-competitive with 1-menaphthyl sulphate, support the suggestion that an Ordered Bi Bi mechanism is operative. 5. Values were determined for some of the dissociation and Michaelis constants for the reaction of the substrates and products with the enzyme. 6. It appears that S-(1-menaphthyl)glutathione activates the enzyme when the concentration of GSH is saturating and that of 1-menaphthyl sulphate is low (of the order of its Michaelis constant).
D062365:Aminobenzoates / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D002850:Chromatography, Gel; D004789:Enzyme Activation; D005978:Glutathione / Q000378:metabolism* / Q000031:analogs & derivatives; D007525:Isoelectric Focusing; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D009281:Naphthalenes / Q000378:metabolism*; D051381:Rats; D013056:Spectrophotometry, Ultraviolet; D013464:Sulfuric Acids / Q000378:metabolism; D014166:Transferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*
D062365:Aminobenzoates; D009281:Naphthalenes; D013464:Sulfuric Acids; C008347:S-(1-menaphthyl)glutathione; D014166:Transferases; D005978:Glutathione