Ceruloplasmin inhibits carbonyl formation in endogenous cell proteins.
Exposure of cells to oxygen radicals results in cellular injury and protein oxidation. Ceruloplasmin is a plasma antioxidant that increases in concentration during inflammation. Therefore, the ability of ceruloplasmin to protect endothelial cells from neutrophil-mediated injury was investigated. The inhibition of protein oxidation by ceruloplasmin was also examined in neutrophil and endothelial cell proteins by analysis of carbonyl formation. In addition, the iron oxidation state was measured to determine the effect of ceruloplasmin ferroxidase activity in oxygen-radical generating systems. Ceruloplasmin significantly (p < .01) inhibited neutrophil-mediated cytotoxicity of endothelial cells by 48%. Carbonyl formation in phorbol myristate acetate (PMA)-stimulated neutrophil proteins was also significantly (p < .01) reduced by ceruloplasmin from 0.172 +/- 0.028 to 0.086 +/- 0.004 mole carbonyl/mole protein. Even though ceruloplasmin itself had a threefold increase in carbonyl formation (0.452 +/- 0.010 vs. 0.146 +/- 0.018 mole carbonyl/mole protein) in the presence of PMA-stimulated compared with unstimulated neutrophils, no loss of functional activity was detected. In xanthine oxidase-treated endothelial cells, ceruloplasmin significantly (p < .05) reduced carbonyl formation from 0.132 +/- 0.010 to 0.097 +/- 0.009 mole carbonyl/mole protein. Ceruloplasmin also significantly (p < .01) oxidized iron when added to PMA-activated neutrophils, thereby decreasing Fe(II) from 98 +/- 8 to 7 +/- 2 microM. Similarly, ceruloplasmin added to xanthine oxidase/hypoxanthine reactions resulted in significant (p < .01) iron oxidation, decreasing Fe(II) from 99 +/- 1 to 15 +/- 3 microM. The ability of ceruloplasmin to protect both endothelial cells and endogenous neutrophil and endothelial cell proteins from oxidative injury suggests that it may be important in regulating cellular and protein damage by oxygen radicals during inflammation.
Krsek-Staples
J A
JA
Department of Internal Medicine, St. Louis University School of Medicine, MO 63110.
Webster
R O
RO
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002417:Cattle; D016923:Cell Death; D002474:Cell-Free System; D002570:Ceruloplasmin / Q000378:metabolism / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000502:physiology*; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D009504:Neutrophils / Q000502:physiology*; D010084:Oxidation-Reduction; D010659:Phenylhydrazines; D011506:Proteins / Q000378:metabolism*; D011651:Pulmonary Artery; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000378:metabolism
D001798:Blood Proteins; D007042:Hypoxanthines; D010659:Phenylhydrazines; D011506:Proteins; C004787:2,4-dinitrophenylhydrazine; D012710:Serum Albumin, Bovine; D019271:Hypoxanthine; D007501:Iron; D002570:Ceruloplasmin; D014969:Xanthine Oxidase; D013755:Tetradecanoylphorbol Acetate