Identification of catechol-protein conjugates in neostriatal slices incubated with [3H]dopamine: impact of ascorbic acid and glutathione.
There is evidence to suggest that degeneration of dopaminergic neurons in Parkinson's disease and certain other conditions results from the action of reactive species generated during the oxidation of dopamine. We, therefore, have begun to explore the conditions under which such reactive species are formed. Tissue slices prepared from rat neostriatum were incubated in a standard Krebs bicarbonate buffer for up to 120 min. In the presence of [3H]dopamine (0.01-100 microM), binding of tritium to the acid-insoluble protein fraction was detected. Binding was attenuated by the addition of ascorbate (0.085-0.85 mM) or glutathione (0.01-1.0 mM) to the buffer. Acid hydrolysis of the protein revealed the presence of cysteinyl-dopamine and cysteinyl-dihydroxyphenylacetic acid residues. These results suggest that dopamine oxidizes to form reactive metabolites, presumably quinones, that then bind to nucleophilic sulfhydryl groups on protein cysteinyl residues. The findings further suggest that the extent to which reactive metabolites are formed is determined in part by the balance between the availability of dopamine and the antioxidant environment.
Hastings
T G
TG
Department of Neuroscience, University of Pittsburgh, Pennsylvania 15260.
Zigmond
M J
MJ
D015102:3,4-Dihydroxyphenylacetic Acid / Q000378:metabolism; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology*; D002396:Catechols / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology*; D008297:Male; D017072:Neostriatum / Q000187:drug effects / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014316:Tritium
D002396:Catechols; D009419:Nerve Tissue Proteins; D014316:Tritium; D015102:3,4-Dihydroxyphenylacetic Acid; D005978:Glutathione; D003545:Cysteine; C034221:catechol; D001205:Ascorbic Acid; D004298:Dopamine