High-sensitivity analysis of sialyl-oligosaccharide glycosylation sites in glycoproteins by miniaturized tryptic digestion and microcolumn liquid chromatography.
A highly sensitive analytical procedure was developed to assess the site of glycosylation in a model glycoprotein, bovine fetuin. First, sample cleavage with immobilized trypsin and the peptide map development are accomplished by microcolumn liquid chromatography. Second, the sialic acids content is measured fluorometrically using their precolumn conversion to quinoxaline derivatives. A unique preconcentration system was developed to secure sensitivity of the second measurement.
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000737:chemistry*; D012794:Sialic Acids / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis / Q000737:chemistry
D006023:Glycoproteins; D009844:Oligosaccharides; D012794:Sialic Acids; D000509:alpha-Fetoproteins; C016735:sialooligosaccharides; D019158:N-Acetylneuraminic Acid