Oxidative modification of human ceruloplasmin by peroxyl radicals.
Ceruloplasmin (CP), the blue oxidase present in all vertebrates, is the major copper-containing protein of plasma. We investigated oxidative modification of human CP by peroxyl radicals generated in a solution containing 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). When CP was incubated with AAPH, the aggregation of proteins was increased in a time- and dose-dependent manner. Incubation of CP with AAPH resulted in a loss of ferroxidase activity. Superoxide dismutase and catalase did not protect the aggregation of CP, whereas hydroxyl radical scavengers such as ethanol and mannitol protected the protein aggregation. The aggregation of proteins was significantly inhibited by the copper chelators, diethyldithiocarbamate and penicillamine. Exposure of CP to AAPH led to the release of copper ions from the enzyme and the generation of protein carbonyl derivatives. Subsequently, when the amino acid composition of CP reacted with AAPH was analyzed, cysteine, tryptophan, methionine, histidine, tyrosine, and lysine residues were particularly sensitive.
Kang
J H
JH
Department of Genetic Engineering, Chongju University, South Korea. jhkang@chongju.ac.kr
Kim
K S
KS
Choi
S Y
SY
Kwon
H Y
HY
Won
M H
MH
D000578:Amidines / Q000737:chemistry; D000596:Amino Acids / Q000032:analysis / Q000737:chemistry; D002374:Catalase / Q000737:chemistry; D002570:Ceruloplasmin / Q000037:antagonists & inhibitors / Q000737:chemistry*; D002614:Chelating Agents / Q000737:chemistry; D003300:Copper / Q000032:analysis; D016166:Free Radical Scavengers / Q000737:chemistry; D006801:Humans; D010084:Oxidation-Reduction; D010545:Peroxides / Q000032:analysis / Q000037:antagonists & inhibitors / Q000737:chemistry*; D012996:Solutions; D013482:Superoxide Dismutase / Q000737:chemistry; D013997:Time Factors
D000578:Amidines; D000596:Amino Acids; D002614:Chelating Agents; D016166:Free Radical Scavengers; D010545:Peroxides; D012996:Solutions; C049375:perhydroxyl radical; C046728:2,2'-azobis(2-amidinopropane); D003300:Copper; D002374:Catalase; D013482:Superoxide Dismutase; D002570:Ceruloplasmin