title: Identification of the thiol groups in human ceruloplasmin.
abstract: Human ceruloplasmin was attached to activated thiol-Sepharose via its thiol groups and was then digested with pepsin. After appropriate washings the thiol peptides were eluted by reduction and were carboxymethylated and purified by column chromatography and electrophoresis. Amino acid sequencing showed that the peptides were derived from five different areas in the molecule and together accounted for 92 residues, six of which were cysteines. Since one of the peptides contained two cysteines it seemed evident that, prior to the reductive elution of the peptides, one of these had been paired in a disulfide bridge with one of the four remaining thiol peptides present in the mixture. The disulfide was isolated and identified by digesting the immobilized protein with pepsin followed by trypsin. The second (tryptic) digestion released the disulfide peptide. Three of the true thiol peptides obtained occur in regions of sequence that have already been reported and which account for 564 of the approximately 1050 residues present in the protein. Three of them also show about 40% identity with each other, whereas no relatedness is observed with the fourth. The three related peptides are, moreover, clearly homologous to the copper-binding areas in the small blue plant and bacterial proteins plastocyanin and azurin. Homologous regions are also evident when the peptides are compared to the two sequences reported for the blue oxidase, fungal laccase, one of which contains a disulfide bridge.
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000302:isolation & purification; D002570:Ceruloplasmin; D055598:Chemical Phenomena; D002621:Chemistry; D004220:Disulfides / Q000302:isolation & purification; D006801:Humans; D010446:Peptide Fragments / Q000302:isolation & purification; D013438:Sulfhydryl Compounds / Q000302:isolation & purification*
chemical_list: D000596:Amino Acids; D004220:Disulfides; D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; D002570:Ceruloplasmin