Separation of limited tryptic fragments of human ceruloplasmin by gel-permeation high-performance liquid chromatography.
Limited tryptic proteolysis of human ceruloplasmin rapidly produces several large, protease-resistant fragments, suggesting that the molecule consists of several domains. In order to locate the sites of proteolytic cleavage in the whole molecule, we used gel-permeation high-performance liquid chromatography to determine the optimum conditions for fragment separation. Using a buffer containing 8 M urea, the 67,000-daltons tryptic fragment from single-chain ceruloplasmin was isolated in a sufficiently pure state for amino acid sequence analysis to determine its location in the uncleaved molecule. These results have been used in conjunction with amino acid sequence data to develop a schematic model of the domain structure of human ceruloplasmin.
D002570:Ceruloplasmin / Q000378:metabolism* / Q000592:standards; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008961:Models, Structural; D010446:Peptide Fragments / Q000302:isolation & purification*; D014357:Trypsin
D010446:Peptide Fragments; D002570:Ceruloplasmin; D014357:Trypsin