Efficient alpha-(alkylthio)alkyl-type radical formation in (*)OH-induced oxidation of alpha-(methylthio)acetamide.

Pulse radiolysis with UV-vis/ESR detection and steady-state gamma-radiolysis, combined with chromatographic techniques, were used to investigate the detailed mechanism of the (*)OH-induced oxidation of alpha-(methylthio)acetamide (alpha-MTA) in aqueous solution. The main pathway involves the formation of hydroxysulfuranyl radicals alpha-MTA-(>S(*)-OH) and alpha-(alkylthio)alkyl radicals H(3)C-S-(*)CH-C( horizontal lineO)-NH(2) (lambda(max) S(*)-OH) radicals undergo efficient conversion to intermolecularly three-electron-bonded dimeric radical cations of alpha-MTA-(>S thereforeS<)(+) (lambda(max) = 480 nm), especially for high alpha-MTA concentrations. In contrast, at low proton concentrations, alpha-MTA-(>S(*)-OH) radicals decompose via the elimination of water, formed through intramolecular hydrogen (attached to the nitrogen atom) transfer to the hydroxysulfuranyl moiety within a six-membered structure. This process leads to the formation of the imine radical H(3)C-S-CH(2)-C( horizontal lineO)(*)NH, which subsequently decays in three independent channels. The first decay channel begins with a beta-scission followed by hydrolysis and a subsequent Hofmann rearrangement. One of the end products of this first decay channel is CO(2), which was detected. The second decay channel involves an intramolecular hydrogen transfer from the deltaC carbon atom to the radical imine site producing the alpha-(alkylthio)alkyl radical H(2)C(*)-S-CH(2)-C( horizontal lineO)-NH(2). In the third decay channel there is a 1,3-hydrogen shift in the imine radical which forms the radical H(3)C-S-(*)CH-C( horizontal lineO)-NH(2). The presence of the amide group induces more complex radical chemistry that leads unexpectedly to the degradation of the CH(3)SCH(2)CONH(2) molecule into gaseous products, CO(2) and NH(3). These features of the mechanism of the (*)OH-induced oxidation of alpha-MTA are quite different from those seen in other organic sulfides in neutral solutions.

Methionine residue acts as a prooxidant in the •OH-induced oxidation of enkephalins.

Enkephalins are bioactive pentapeptides (Tyr-Gly-Gly-Phe-Leu (Leu-enk) and Tyr-Gly-Gly-Phe-Met (Met-enk)) produced while an organism is under mental and/or physical stress. In the course of their biological action they are exposed to reactive oxygen and nitrogen species. We have reinvestigated the reactions of (•)OH radicals toward these peptides in order to elucidate the oxidation mechanisms and the final products. Nanosecond pulse radiolysis was used to obtain the spectra of the reaction intermediates and their kinetics. Additional insight into details of the oxidation mechanism was gained by identification of main final products by means of UV-vis spectrophotometry, HPLC coupled with fluorescence spectroscopy, and mass spectrometry. The key processes are different in both peptides. In Leu-enk, the first step is an (•)OH radical addition to the aromatic rings of Tyr and Phe residues that leads to hydroxylated residues, dihydroxyphenylalanine (DOPA) from Tyr and tyrosine isomers from Phe, respectively. In Met-enk, these processes are less important, an additional target being the sulfur atom of the methionine residue. Depending on pH either an OH-adduct (hydroxysulfuranyl radical) or a sulfur radical cation undergo intramolecular electron transfer with Tyr residue resulting in a repair of Met and oxidation of Tyr to tyrosyl radicals and a final formation of dityrosine. At low pH, the OH-adducts to Tyr residue are precursors of tyrosyl radicals and dityrosine. Thus, the final products coming from oxidation of the Tyr residue depend strongly on the neighboring residues and the pH.