thiamine-disulfide and oxodihydrothiochrome

It is shown that nitrogen dioxide oxidizes thiamine to thiamine disulfide, thiochrome, and oxodihydrothiochrome (ODTch). The latter is formed during oxidation of thiochrome by nitrogen dioxide. Nitrogen dioxide was produced by incubation of nitrite with horse ferric myoglobin and human hemoglobin in the presence of hydrogen peroxide. After addition of tyrosine or phenol to aqueous solutions containing oxoferryl forms of the hemoproteins, thiamine, and nitrite, the yield of thiochrome greatly increased, whereas the yield of ODTch decreased. In the presence of high concentrations of tyrosine or phenol compounds ODTch was not formed at all. The neutral form of thiamine with the closed thiazole cycle and minor tricyclic form of thiamine do not enter the heme pocket of the protein and do not interact with the oxoferryl heme complex Fe(IV=O) or porphyrin radical. The tricyclic form of thiamine is oxidized to thiochrome by tyrosyl radicals located on the surface of the hemoprotein. The thiol form of thiamine is oxidized to thiamine disulfide by both hemoprotein tyrosyl radicals and oxoferryl heme complexes. Nitrite and also tyrosine, tyramine, and phenol readily penetrate into the heme pocket of the protein and reduce the oxyferryl complex to ferric cation. These reactions yield nitrogen dioxide as well as tyrosyl and phenoxyl radicals of tyrosine molecules and phenol compounds, respectively. Tyrosyl and phenoxyl radicals of low molecular weight compounds oxidize thiamine only to thiochrome and thiamine disulfide. The effect of oxoferryl forms of myoglobin and hemoglobin, nitrogen dioxide, and phenol on thiamine oxidative transformation as well as antioxidant properties of the hydrophobic thiamine metabolites thiochrome and ODTch are discussed.

Topics: Animals; Hemoglobins; Horses; Humans; Hydrogen Peroxide; Kinetics; Myoglobin; Nitrites; Nitrogen Dioxide; Oxidation-Reduction; Thiamine

Effects of thiamine and its derivatives on inhibition of dityrosine formation were studied in reactions catalyzed by oxoferryl forms of hemoglobin. At high thiamine concentrations, a complete inhibition of dityrosine formation was observed due to interaction of tyrosyl radicals with thiamine tricyclic and thiol forms. In neutral and alkaline media, tyrosyl radicals oxidized thiamine to thiochrome, oxodihydrothiochrome, and thiamine disulfide. In the absence of tyrosine, oxoferryl forms of hemoglobin manifested peroxidase activity towards thiamine and its phosphate esters by inducing their oxidation to disulfide compounds, thiochrome, oxodihydrothiochrome, and their phosphate esters, respectively, in neutral media. Thiamine and its phosphate esters were oxidized by both oxoferryl forms of hemoglobin, viz., +*Hb(IV=O) (compound I with an additional radical on the globin) and Hb(IV=O) (compound II). Putative mechanisms of thiamine conversions under oxidative stress and the protective role of hydrophobic thiamine metabolites are discussed.

Topics: Hemoglobins; Humans; Kinetics; Metmyoglobin; Oxidation-Reduction; Peroxidase; Thiamine; Tyrosine

After peroxynitrite addition to aqueous solutions of thiamine at neutral and alkaline pH formation of thiamine disulfide and fluorescent products was observed. The fluorescent compounds were identified as thiochrome (TChr) and oxodihydrothiochrome (ODTChr) using spectral and fluorescent methods as well as paper chromatography and mass spectrometry. TChr and ODTChr are not the end products of thiamine oxidation and in neutral medium are unstable to peroxynitrite action and degrade rapidly to form non-fluorescent products. Thiamine, TChr, and ODTChr protects tyrosine from its modification by peroxynitrite. In the presence of TChr and ODTChr modification of tyrosinyl residues in human serum albumin and cytocrome c decreased. The prolonged thiamine incubation with glucose, amino acids and nitrite was accompanied by oxidative transformation of thiamine and formation of fluorescent products. We have shown that thiamine is also oxidized into TChr and ODTChr, i.e., it forms the same products as after thiamine oxidation by peroxynitrite. Moreover, thiamine (or its derivatives) appears as peroxynitrite scavenger leading to toxic effects lowering at diabetes mellitus.

Topics: Diabetic Angiopathies; Humans; Hydrogen-Ion Concentration; Models, Biological; Oxidation-Reduction; Peroxynitrous Acid; Proteins; Thiamine