Analysis of chicken and turkey ovalbumins by microchip electrophoresis combined with exoglycosidase digestion.
The polypeptide and carbohydrate patterns of two glycoproteins, chicken ovalbumin (CO) and turkey ovalbumin (TO), were analyzed by microchip electrophoresis (ME), following digestion with proteases and exoglycosidases. Glycopeptides derived from ovalbumin were obtained by digestion with Pronase, followed by dialysis, and then separated by ME. Using CO as model, the method was developed to deduce the structure of glycans from glycoproteins by comparing the electropherograms of glycopeptides with and without digestion of exolycosidases. Applying the same approach, the structure of oligosaccharides linked to TO was determined. TO was found to contain high-mannose type oligosaccharides and oligosaccharides with terminal N-acetylglucosamine residues. The complete primary analysis of CO and TO by ME described in this paper provides a basis for an analysis of glycoproteins with an integrated microfluidic chip.
D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D002645:Chickens; D004586:Electrophoresis / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis / Q000737:chemistry; D006026:Glycoside Hydrolases; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000032:analysis* / Q000737:chemistry; D010449:Peptide Mapping; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry; D011402:Pronase; D040081:Protein Array Analysis; D014357:Trypsin; D014421:Turkey
D006020:Glycopeptides; D011134:Polysaccharides; D010047:Ovalbumin; D006026:Glycoside Hydrolases; D014357:Trypsin; D011402:Pronase