Photoinactivation of agaricus bisporus tyrosinase: modification of the binuclear copper site.
Irradiation of Agaricus bisporus tyrosinase in the presence of citrate at pH 5.6 with 300-420-nm light results in a loss of both catecholase activity and cresolase activity. The light-sensitive species appears to be an enzyme-citrate complex, most likely involving coordination of citrate to the active site copper. One copper ion from each binuclear active site can be removed from the inactivated enzyme, resulting in the formation of a met apo derivative. The electron spin resonance spectrum of met apo tyrosinase resembles that of met apo hemocyanin and half-met Neurospora tyrosinase. It is consistent with a distorted square-planar geometry around the copper and with either nitrogen or nitrogen and oxygen ligands. Amino acid analysis indicates that four histidines on the heavy subunit are destroyed during the inactivation process. Some or all of these histidines may serve as ligands to the copper ion which becomes labile after inactivation. Photoinactivation results in decarboxylation of citrate and does not require the presence of oxygen. The reaction may involve generation of a free radical from the citrate which then attacks nearby histidine residues.
D000363:Agaricales / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D004156:Catechol Oxidase / Q000528:radiation effects*; D003300:Copper / Q000378:metabolism*; D007700:Kinetics; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors / Q000528:radiation effects*; D010782:Photolysis; D014466:Ultraviolet Rays
D000596:Amino Acids; D003300:Copper; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase