A rat brain fraction and different purified peroxidases catalyzing the formation of dopaminochrome from dopamine.
Dopaminochrome formation is catalyzed by commercially available purified peroxidases (EC 1.11.1.7) such as horseradish, lacto- and myelo-peroxidase using dopamine, hydrogen peroxide or promethazine sulfoxide as substrates. A rat brain fraction (RBF) catalyzes a similar reaction and its catalytic power increases after preincubation with hydrogen peroxide/ascorbic acid. The activity of both the purified enzymes and the RBF preparation is inhibited by carnosine and characterized by excess substrate inhibition. The enzymes recognize different substrates but show the highest affinity for dopamine. The RBF fraction is strongly buffered against oxidation by compounds such as glutathione and by bioreductive enzymes such as DT-diaphorase (EC 1.6.99.2) which can use as a substrate menadione or dopaminochrome. The rat brain dopamine peroxidizing activity appeared to be mostly bound to the synaptosomal fraction. The reaction catalyzed by the purified peroxidases was followed by electron spin resonance spectroscopy and, unlike that catalyzed by RBF, was shown to produce the signal of a transient dopamine-o-semiquinone radical.
Galzigna
L
L
Department of Biological Chemistry, University of Padua, viale G. Colombo 3, 35121, Padua, Italy. gal@civ.bio.unipd.it
Schiappelli
M P
MP
Rigo
A
A
Scarpa
M
M
D000818:Animals; D001921:Brain / Q000378:metabolism*; D004298:Dopamine / Q000737:chemistry / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D006863:Hydrogen-Ion Concentration; D045563:Indolequinones; D007211:Indoles / Q000737:chemistry / Q000378:metabolism*; D007784:Lactoperoxidase / Q000378:metabolism; D008297:Male; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013574:Synaptosomes / Q000378:metabolism
D045563:Indolequinones; D007211:Indoles; C091584:aminochrome 1; D007784:Lactoperoxidase; D010544:Peroxidases; D004298:Dopamine