Amino acids side-chain peroxidation

Electron-transfer reactions between cytochrome c and cytochrome c peroxidase have been studied extensively because of the well-characterized structures and biophysical properties of the reactants [146-150]. It is well known that the resting ferric form of cytochrome c peroxidase is oxidized by hydrogen peroxide to compound I, which contains an oxyferryl heme moiety in which the iron atom has a formal oxidation state of 4-1- [146-150]. The other is a porphyrin n radical cation or organic radical (R +) localized on an amino acid residue of Trp-191 [151-154] this is formed by transfer of the oxidized equivalent to the amino acid side chain [150]. The site of electron transfer in the reduction of compound I has been controversial and two forms of compound II have been identified, (P)Fe =0 containing the oxyferryl heme Fe(IV) [155-158] and [(P)Fe ] + containing Fe(III) and the porphyrin % radical cation which oxidizes the amino acid side-chain to produce an organic radical [(P)Fe +, R" ] [159 165] as shown in Scheme 10. 

Oxidative amino acid side-chain modifications do not result in a stable end product of the oxidation process, but very often highly reactive intermediates are formed. These include chemically reactive groups, like ketones and aldehydes, or the formation of protein hydroperoxides. The presence of such protein hydroperoxides leads to a process called protein peroxidation. Here secondary reactions occur if the protein hydroperoxide decomposes and initiates further oxidative reactions, again forming oxidized protein forms. 

Oxidation occurs generally on the amino acid side chains due to exposure to air, residual peroxide from excipients, or exposure to visible or ultraviolet light. In particular, methionine, cysteine, tryptophan, and tyrosine are prone to oxidation. Metal ions such as iron, zinc, copper, or tungsten from metals that are used in the manufacturing process, leached from contact materials, or present in trace amounts in excipients can catalyze oxidation as well as other degradation processes [2, 17]. 

Hydroxyl radical oxidative modification, one of the footprinting approaches, utilizes hydroxyl radicals to oxidize certain amino acid side chains followed by extent of modification measurement by MS. For example, fast photochemical oxidation of proteins (FPOP) [35] involves irradiation of protein sample solutions containing hydrogen peroxide by a KrF excimer laser and generation of hydroxyl radicals in the solution. FPOP occurs on the microsecond timescale for the labeling... 

Amino acid residues are potential targets of free radical oxidation and nitration. Carbonyl derivatives of proteins may be formed by the interaction of protein amino acid side chains, mainly cysteine, histidine, and lysine residues with reactive aldehydes, such as HNE and ONE generated by peroxidation of PUFAs (polyunsaturated fatty acids). Amino acid and peptide biomarkers of oxidative stress are typically focused on specific proteins related to disease pathology. For instance, the oxidation of histidine and methionine are typically discussed in (3-amyloid plaque formation and HNE-derived histidine adducts are the main focus of modifications on low-density lipoprotein (LDL) (An-nangudi et al., 2008). However, there are several specific examples of general biomarkers of oxidative stress that include endogenous histidine containing dipeptides such as carnosine and anserine as well as the very stable o,o -dityrosine. These will be discussed below. 

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