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Biomedical subjects

P J O'Brien

Publications and source records attributed to P J O'Brien.

At least 325 records · Page 18Linked to original sources
PubMed1968

The effectiveness of a lipid peroxide in oxidizing protein and non-protein thiols.

1. Thiol oxidation by a lipid peroxide or hydrogen peroxide was as efficient in denatured non-haem proteins as in small thiols. Both peroxides were relatively ineffective in oxidizing haemoprotein thiols, especially at low pH. Increased amounts of haematin decreased greatly the efficiency of GSH oxidation by peroxides especially at low pH. 2. Other than the haematin ring, the thiol group was found to be probably the group in proteins most sensitive to modification by peroxides. 3. At low concentrations, the fatty acid moiety of a lipid peroxide appeared to impede thiol oxidation in proteins, probably by hydrophobic bonding to the protein, rather than to stimulate thiol oxidation by denaturing the protein and thereby increasing the exposure and reactivity of the thiol group. 4. The relative rates of thiol oxidation by peroxides in the different thiols were: haemoprotein thiols>small thiols>other protein thiols. In all cases, thiol oxidation was much more rapid by the lipid peroxide than by hydrogen peroxide.

Adult↗
PubMed1967

Xanthine oxidase inactivation by reagents that modify thiol groups.

1. The presence of xanthine was required for the inhibition of bovine milk xanthine oxidase by o-iodosobenzoate, iodoacetamide, hydrogen peroxide or p-chloromercuribenzoate. 2. Inactivation by p-chloromercuribenzoate was very rapid, was reversed by cysteine and was less in the presence of FAD. Lineweaver-Burk plots showed that the inactivation by p-chloromercuribenzoate was competitive with substrate. 3. Inactivation by o-iodosobenzoate, iodoacetamide or hydrogen peroxide could not be reversed by cysteine or xanthine. However, the presence of xanthine during the incubation with inhibitor protected the enzyme against o-iodosobenzoate but not against iodoacetamide or hydrogen peroxide. 4. p-Chloromercuribenzoate protected the enzyme against inactivation by hydrogen peroxide.

Amides↗
PubMedDate not supplied

Cytochrome P450 dependent xenobiotic activation by physiological hydroperoxides in intact hepatocytes.

Xenobiotic metabolic activation by intact hepatocytes was recently shown to be enhanced by the addition of nontoxic concentrations of t-butyl hydroperoxide and prevented by cytochrome P450 inhibitors. Furthermore, H2O2 (Km = 103 microM) was found to be highly effective in supporting the human microsomal CYP1A2 catalyzed metabolic activation of the heterocyclic aromatic amine 2-amino-3-methylimidazo (4,5-f) quinoline (IQ) to mutagenic metabolites and the DNA adduct formed was the same as that formed by the mixed-function oxidase catalyzed activation system. In the following, it is shown that the cytotoxicity of other xenobiotics including carcinogenic arylamines and their N-hydroxyarylamine metabolites were markedly enhanced by hydroperoxide addition but not in the presence of cytochrome P450 inhibitors. The CYP1A2 dependent O-demethylation of methoxyresorufin in 3-methylcholanthrene induced hepatocytes was also markedly enhanced when intracellular H2O2 was generated by the mitochondrial monoamine oxidase (MAO) substrates tyramine or kynurenamine. Linoleic acid hydroperoxide also dramatically enhanced the cytotoxicity of phenelzine towards isolated hepatocytes and the microsomal metabolism of phenelzine to form ethylbenzene. The P450 inhibitors phenylimidazole, benzylimidazole prevented the metabolic activation of phenelzine but not lipid peroxidation. These results suggest that linoleic acid hydroperoxide can activate hydrazines via a cytochrome P450 peroxidase catalyzed one electron oxidation to form highly cytotoxic reactive intermediates. Furthermore, increased hydrogen peroxide formation, e.g. as a result of oxidative stress, would also be expected to enhance the metabolic activation of carcinogenic arylamines via the peroxygenase function of CYP1A2.

Animals↗
PubMedDate not supplied

Peroxidase-mediated irreversible binding of arylamine carcinogens to DNA in intact polymorphonuclear leukocytes activated by a tumor promoter.

Addition of the tumor promoter phorbol myristate acetate to polymorphonuclear leukocytes results in the oxidation of the arylamine carcinogens; [14C]benzidine, N-[14C]methylaminoazobenzene and [14C]aminofluorene to reactive intermediate(s) that bind irreversibly to the leukocyte DNA. The binding was dependent on oxygen and was decreased by sulfhydryl inhibitors and phenolic antioxidants that inhibit the respiratory burst triggered by the phorbol myristate. Both the binding and the respiratory burst were increased by azide, presumably as a result of intracellular catalase inhibition. However higher concentrations of azide and cyanide prevented binding without affecting the respiratory burst indicating that myeloperoxidase is a catalyst for the binding. Granules isolated from the activated leukocytes and H2O2 catalyzed a cyanide sensitive benzidine binding to calf thymus DNA. Myeloperoxidase and H2O2 also catalysed extensive binding of these arylamines to calf thymus DNA. The leukocytes appear to be a useful model cell for studying one electron oxidation-catalyzed carcinogen activation.

Amines↗