Purification of glycopeptides of human plasma proteins by high-performance liquid chromatography.
The combination of gel permeation chromatography and high-performance liquid chromatography proves to be very effective for the purification of high-molecular-weight glycopeptides containing a single glycan, that have been difficult to separate by other procedures. In order to facilitate comparison of the chromatographic properties of glycopeptides derived from a variety of proteins and having different structures, identical procedures were used for their purification. The method was applied to a series of human plasma proteins, including immunoglobulin D, ceruloplasmin, hemopexin, beta-2-glycoprotein I, 3.1S alpha-2-leucine-rich glycoprotein, and alpha-1-B-glycoprotein. All the purified glycopeptides were placed in the protein structure of these plasma proteins. In several cases the carbohydrate structure has been determined by collaborating groups. Immunoglobulin D is the first example of a glycoprotein whose entire primary structure has been defined by utilizing a a single protein source. Furthermore, hemopexin and 3.1S alpha-2-leucine-rich glycoprotein were both found to contain GalN oligosaccharide, which had not previously been identified in these proteins. The method was also used to identify the oligosaccharide that is missing in a carbohydrate variant of ceruloplasmin.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001798:Blood Proteins / Q000302:isolation & purification*; D002241:Carbohydrates / Q000032:analysis; D002570:Ceruloplasmin / Q000032:analysis; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006020:Glycopeptides / Q000302:isolation & purification*; D006801:Humans; D006868:Hydrolysis; D007072:Immunoglobulin D / Q000032:analysis; D009844:Oligosaccharides / Q000032:analysis
D000596:Amino Acids; D001798:Blood Proteins; D002241:Carbohydrates; D006020:Glycopeptides; D007072:Immunoglobulin D; D009844:Oligosaccharides; D002570:Ceruloplasmin