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Direct evidence of ceruloplasmin antioxidant properties

Molecular and Cellular Biochemistry 1998

Abstract

The chain-breaking antioxidant potential of caeruloplasmin and bovine serum albumin (BSA) has been investigated in comparisonwith other well-established antioxidants. Their Oxygen Radical Absorbing Capacity (ORAC), was measured by usingß-phycocyanin (ß-PC) as a fluorescent indicator protein, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) as a peroxyl radicalgenerator and the water soluble vitamin E analogue, Trolox, as a reference standard. The relative peroxyl absorbing capacities/mole for Trolox, caeruloplasmin, heat-denatured caeruloplasmin (hCP), catalase, bovine serum albumin (BSA), superoxidedismutase (SOD), and deferoxamine were 1; 2.6; 3.3; 3.7; 1.2; 0.1; 0.2, respectively. Caeruloplasmin was far more effective asa peroxyl radical scavenger than SOD, deferoxamine and BSA, but slightly less effective than catalase. The peroxyl radicalabsorbing capacity of caeruloplasmin was enhanced by heat-denaturation of the protein. Electron paramagnetic resonance (EPR)spectroscopy using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin-trap, was applied in order to measure the scavenger abilitiesof caeruloplasmin on superoxide radical and hydroxyl radical production and the concentration required to inhibit by 50% oxygenfree radical formation (IC50) was determined. The IC50 values of caeruloplasmin, hCP, and BSA for the superoxide radical were12, 2, 260 µM and for the hydroxyl radical 15, 2, 200 µM. These results show that caeruloplasmin is an effective chain-breakingantioxidant for a variety of radicals, independently of its catalytic ferroxidase activity. (
1998AbstractThe chain-breaking antioxidant potential of caeruloplasmin and bovine serum albumin (BSA) has been investigated in comparisonwith other well-established antioxidants. Their Oxygen Radical Absorbing Capacity (ORAC), was measured by usingß-phycocyanin (ß-PC) as a fluorescent indicator protein, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) as a peroxyl radicalgenerator and the water soluble vitamin E analogue, Trolox, as a reference standard. The relative peroxyl absorbing capacities/mole for Trolox, caeruloplasmin, heat-denatured caeruloplasmin (hCP), catalase, bovine serum albumin (BSA), superoxidedismutase (SOD), and deferoxamine were 1; 2.6; 3.3; 3.7; 1.2; 0.1; 0.2, respectively. Caeruloplasmin was far more effective asa peroxyl radical scavenger than SOD, deferoxamine and BSA, but slightly less effective than catalase. The peroxyl radicalabsorbing capacity of caeruloplasmin was enhanced by heat-denaturation of the protein. Electron paramagnetic resonance (EPR)spectroscopy using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin-trap, was applied in order to measure the scavenger abilitiesof caeruloplasmin on superoxide radical and hydroxyl radical production and the concentration required to inhibit by 50% oxygenfree radical formation (IC50) was determined. The IC50 values of caeruloplasmin, hCP, and BSA for the superoxide radical were12, 2, 260 µM and for the hydroxyl radical 15, 2, 200 µM. These results show that caeruloplasmin is an effective chain-breakingantioxidant for a variety of radicals, independently of its catalytic ferroxidase activity.

Full paper https://www.researchgate.net/publication/226510548_Direct_evidence_of_ceruloplasmin_antioxidant_properties