Purification and partial characterization of goose ceruloplasmin.
The preparation and properties of ceruloplasmin from goose blood plasma are described. Ammonium sulfate was used to precipitate the crude protein followed by adsorption and elution from DEAE-Sephadex A-50. Further treatment with an ethanol-chloroform mixture and Sephadex G-200 yielded an intensely blue protein possessing a high degree of chemical purity and biological activity. Goose ceruloplasmin, existing in two forms, appears to be a single polypeptide, apparent Mr121,300, with an A610/A280 ratio of 0.07. Copper represented 0.32%, which corresponded to six atoms of copper per protein molecule. Although the amount of EPR-detectable copper was the same as in mammalian ceruloplasmins there were some differences in EPR parameters, mainly in A parallel. Goose ceruloplasmin's amino acid composition, although similar in many residues to human ceruloplasmin, was lower in tyrosine, cystine/cysteine, and acidic amino acids. Valine was found as the N-terminal amino acid. Hexose, hexosamine, sialic acid, and fucose accounted for 6.65% of the weight. Goose protein contained only half the sialic acid of human ceruloplasmin. Two values for Km using either p-phenylenediamine (0.64 and 0.053 mM) or o-dianisidine (0.76 and 0.15 mM) were evaluated from Lineweaver-Burk plots. EPR studies on reactions with water radiolysis products at cryogenic temperatures allowed us to discover that goose ceruloplasmin, like human and bovine ceruloplasmins, possesses superoxide dismutase activity.
Hilewicz-Grabska
M
M
Institute of Biochemistry, University of Lodz, Poland.
Zgirski
A
A
Krajewski
T
T
Plonka
A
A
D000596:Amino Acids / Q000097:blood; D000818:Animals; D002241:Carbohydrates / Q000097:blood; D002570:Ceruloplasmin / Q000302:isolation & purification*; D003300:Copper / Q000097:blood; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D005777:Geese / Q000097:blood*; D007527:Isoenzymes / Q000097:blood; D008970:Molecular Weight; D010088:Oxidoreductases / Q000097:blood; D013481:Superoxides
D000596:Amino Acids; D002241:Carbohydrates; D005609:Free Radicals; D007527:Isoenzymes; D013481:Superoxides; D003300:Copper; D010088:Oxidoreductases; D002570:Ceruloplasmin