Isolation and fractionation of glycopeptides from porcine thyroglobulin.
1. Glycopeptides were isolated by gel filtration on Sephadex G-25 and Sephadex G-50 from a Pronase digest of porcine thyroglobulin. 2. Isolated glycopeptides were separated into five main fractions on a column of DEAE-Sephadex A-25. Of these fractions I to III were further purified by SE-Sephadex C-25 or DEAE-Sephadex A-25 column chromatography. Several of the purified glycopeptides were homogeneous on paper electrophoresis. 3. Based on the chemical composition and molecular weight of the fractionated glycopeptides, two distinct types of heterosaccharide chain were demonstrated. 4. One type of the heterosaccharide unit consisted of four to eight residues of mannose and two residues of glucosamine and had a molecular weight of 1000-1700. The other type of unit contained sialic acid, fucose and galactose in addition to mannose and glucosamine and had a molecular weight of about 3600. 5. Mild alkaline treatment of the glycopeptide did not result in the destruction of threonine and serine. 2-Acetamido-1-N-(4-l-aspartyl)-2-deoxy-beta-d-glucopyranosylamine was isolated from partial acid hydrolysates.
D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004586:Electrophoresis; D005643:Fucose / Q000032:analysis; D005690:Galactose / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D006020:Glycopeptides / Q000302:isolation & purification; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight; D009438:Neuraminic Acids / Q000032:analysis; D010455:Peptides / Q000302:isolation & purification*; D011134:Polysaccharides / Q000302:isolation & purification*; D011402:Pronase; D012694:Serine / Q000032:analysis; D013552:Swine; D013912:Threonine / Q000032:analysis; D013954:Thyroglobulin / Q000032:analysis*
D006020:Glycopeptides; D009438:Neuraminic Acids; D010455:Peptides; D011134:Polysaccharides; D005643:Fucose; D013912:Threonine; D012694:Serine; D013954:Thyroglobulin; D011402:Pronase; D005944:Glucosamine; D008358:Mannose; D005690:Galactose