Preparation of cyanogen bromide fragments of MM, NN, and MN glycoproteins (glycophorins) from human erythrocyte membranes of single donors.
A procedure is described for the preparation of three cyanogen bromide fragments of the MM, NN, or MN glycoprotein (glycophorin) of the human erythrocyte membranes, from erythrocytes of single donors. The fragments are obtained in pure form and excellent yields by employing procedures which include proteolytic inhibitors during membrane processing, thorough delipidation of the glycoprotein, and CNBr cleavage conditions which lead to quantitative fragmentation without loss of carbohydrates. A phenol-urea extraction resolves the two glycopeptide fragments from the carbohydrate-free fragment. The two glycopeptides are further purified by Bio-Gel P-6 and P-100 chromatography. The three fragments include the amino terminal 8 residue glycopeptide, a large glycopeptide form the middle of the molecule which bears the Asn-linked oligosaccharide and 8--9 O-glycosidically linked units, and a carboxyl terminal, carbohydrate-free, approx. 50 residue fragment. Their amino acid and carbohydrate composition, and size, are in close agreement with the sequence data of Tomita, M., Furthmayr, H. and Marchesi, V.T. (Biochemistry (1978), 17, 4756--4770). The fragments represent three well delineated portions of the glycoprotein molecule.
D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis*; D004912:Erythrocytes / Q000032:analysis*; D006020:Glycopeptides / Q000032:analysis; D006021:Glycophorins / Q000302:isolation & purification; D006801:Humans; D008951:MNSs Blood-Group System; D010446:Peptide Fragments / Q000032:analysis; D012795:Sialoglycoproteins / Q000302:isolation & purification
D000596:Amino Acids; D002241:Carbohydrates; D006020:Glycopeptides; D006021:Glycophorins; D008951:MNSs Blood-Group System; D010446:Peptide Fragments; D012795:Sialoglycoproteins; D003488:Cyanogen Bromide