Abstract
Using the fluorescent amino acid tryptophan (Trp), we have characterized the copper(II) binding of F4W alpha-synuclein in the presence and absence of dioxygen at neutral pH. Variations in Trp fluorescence indicate that copper(II) binding is enhanced by the presence of dioxygen, with the apparent dissociation constant (K(d(app))) changing from 100nM (anaerobic) to 10nM (aerobic). To investigate the possible role of methionine oxidation, complementary work focused on synthetic peptide models of the N-terminal Cu(II)-alpha-syn site, MDV(F/W) and M( *)DV(F/W), where M( *)=methionine sulfoxide. Furthermore, we employed circular dichroism (CD) spectroscopy to demonstrate that the phenyl-to-indole (F-->W) substitution does not alter copper(II) binding properties and to confirm the 1:1 metal-peptide binding stoichiometry. CD comparisons also revealed that Met1 oxidation does not affect the copper-peptide conformation and further suggested the possible existence of a Cu(II)-Trp/Phe (cation-pi) interaction.