Neuronal Proteins as Targets of 3-Hydroxykynurenine: Implications in Neurodegenerative Diseases

The neurotoxic activity of the tryptophan metabolite 3-hydroxykynurenine (3OHKyn) in neurodegenerative disorders, such as Parkinson's and Alzheimer's diseases, is related to oxidative stress and 3OHKyn interaction with cellular proteins. The pattern of protein modification induced by 3OHKyn involves the nucleophilic side chains of Cys, His, and Lys residues, similarly to the one promoted by dopamine and other catecholamines. In the present work, we have analyzed the reactivity of 3OHKyn toward the neuronal targets alpha-synuclein (and its N-terminal fragments 1-6 and 1-15) and amyloid-beta peptides (1-16 and 1-28) and characterized the resulting conjugates through spectrometric (LCMS/MS) and spectroscopic (UV-vis, fluorescence, NMR) techniques. The amino acid residues of alpha-synuclein and amyloid-beta peptides involved in derivatizations by 3OHKyn and its autoxidation products (belonging to the xanthommatin family) are Lys and His, respectively. The pattern of protein modification is expanded in the conjugates obtained in the presence of the metal ions copper(II) or iron(III), reflecting a more oxidizing environment that in addition to adducts with protein/peptide residues also favors the fragmentation of the protein. These results open the perspective to using the 3OHKyn-protein/peptide synthetic conjugates to explore their competence to activate microglia cell cultures as well as to unravel their role in neuroinflammatory conditions.