title: Purification and partial characterization of goose ceruloplasmin.
abstract: 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.
authors.affiliation: Institute of Biochemistry, University of Lodz, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Hilewicz-Grabska
authors.forename: A
authors.initials: A
authors.lastname: Zgirski
authors.forename: T
authors.initials: T
authors.lastname: Krajewski
authors.forename: A
authors.initials: A
authors.lastname: Plonka
mesh_terms: 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
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D005609:Free Radicals; D007527:Isoenzymes; D013481:Superoxides; D003300:Copper; D010088:Oxidoreductases; D002570:Ceruloplasmin