Date of Degree
PhD (Doctor of Philosophy)
Jonathan A. Doorn
Parkinson's disease (PD) is a progressive neurodegenerative and movement disorder that involves specific loss of dopaminergic neurons in the substantia nigra of the brain. Exact causes of PD are unknown. However, cells affected in PD are centers of dopamine (DA) synthesis, storage, and metabolism, which implicate DA as an endogenous neurotoxin that contributes to PD. Furthermore, DA is known to undergo oxidation to radicals and quinones. These reactive species exert deleterious effects on cells through a variety of mechanisms that are relevant to the pathogenesis of PD. Another potential mechanism of toxicity for DA is metabolism to 3,4-dihydroxyphenylacetaldehyde (DOPAL). This reactive metabolite is significantly more toxic than the parent DA. DOPAL has several demonstrated mechanisms of toxicity, including formation of protein-adducts via reaction with amine-type cellular nucleophiles. However, known toxicity mechanisms do not fully account for DOPAL's high toxicity. Oxidation of DOPAL to a reactive quinone or radical could help explain its high toxicity. Therefore, the hypothesis of this work is that DOPAL is capable of undergoing oxidation that leads to increased protein modification and nucleophilic reactivity. Experimentally, oxidation of DOPAL results in formation of a semi-quinone radical and an ortho-quinone, as confirmed by electron paramagnetic resonance spectroscopy and nuclear magnetic resonance spectroscopy, respectively. In agreement with the stated hypothesis, oxidation of DOPAL enhanced its ability to induce protein cross-linking of a model protein (glyceraldehyde 3-phosphate dehydrogenase) as indicated by polyacrylamide gel-electrophoresis. Also, the presence of anti-oxidants (ascorbate, N-acetyl cysteine) attenuated the reactivity of DOPAL with the model aminenucleophile N-acetyl lysine. These results indicate that DOPAL oxidation enhances both protein cross-linking and nucleophilic reactivity.
This work resulted in several other important findings. DOPAL is shown to undergo carbonyl-hydration in aqueous media, and spontaneous oxidation of DOPAL results in formation of superoxide. Furthermore, DOPAL is shown to be susceptible to oxidation by cyclooxygenase-2, an enzyme known to be involved in PD. This provides a potential mechanism for formation of the oxidized products identified here. As DA metabolism and oxidation occur in cells affected by PD, the experimental results demonstrated here are likely relevant for understanding the pathogenesis of PD.
3,4-Dihydroxyphenylacetaldehyde, Dopamine, Dopamine Metabolism, Oxidation, Parkinson's Disease, Reactive Intermediates
xviii, 163 pages
Includes bibliographical references (pages 150-163).
Copyright 2011 David Anderson