Structure of a major oligosaccharide of PASII/PMP22 glycoprotein in bovine peripheral nerve myelin.
The amino acid sequence of the glycopeptide obtained from bovine PASII/PMP22 protein in the PNS myelin was determined to be Gln-Asn-Cys-Ser-Thr, where the asparagine was glycosylated. To eliminate all the contaminated P(o) glycopeptides from the PASII/PMP22 glycopeptide preparation, we used a fluorescent probe, N-[2-(2-pyridylamino)ethyl]maleimide, which reacts with the cysteine of the PASII/PMP22 glycopeptides. The labeled PASII/PMP22 glycopeptides were isolated by HPLC and were digested further with glycopeptidase A. The resultant oligosaccharides were conjugated with 2-aminopyridine (PA) as a fluorescent tag. One major PA-oligosaccharide, OPPE1, was purified by HPLC. The structure of OPPE1 was elucidated by fast atom bombardment mass spectrometry and (1)H-NMR studies and comparing the derivatives of PA-OPPE1 and PA-oligosaccharides of gamma-globulin on HPLC. The structure, SO(4)-3GlcAbeta1-3Galbeta1-4GlcNAcbeta1-2Manalpha1+ ++-6(GlcNAcbeta1-4) (GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1- 4(Fucalpha1-6)GlcNAc- PA, was identical to the pyridylaminated form of the major oligosaccharide D8 of bovine P(o) previously reported.
Kitamura
K
K
Department of Physiology, Saitama Medical School, Saitama, Japan. Department of Physiology, Faculty of Medicine, Keio University, Tokyo, Japan. kitamura@saitamamed.ac.jp
Uyemura
K
K
Shibuya
K
K
Sakamoto
Y
Y
Yoshimura
K
K
Nomura
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006023:Glycoproteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000737:chemistry*; D009186:Myelin Sheath / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry; D010525:Peripheral Nerves / Q000737:chemistry*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D006023:Glycoproteins; D009185:Myelin Proteins; D009844:Oligosaccharides; D010446:Peptide Fragments