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 Po glycopeptides from the PASII/PMP22 glycopeptide preparation, we used a fluoresc...

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Published in:Journal of neurochemistry 2000-08, Vol.75 (2), p.853-860
Main Authors: Kitamura, Kunio, Uyemura, Keiichi, Shibuya, Kyoko, Sakamoto, Yasushi, Yoshimura, Kazunori, Nomura, Masahiko
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Animals
Asparagine‐linked oligosaccharide
Biological and medical sciences
Carbohydrate Conformation
Carbohydrate Sequence
Cattle
Chromatography, High Pressure Liquid
Fundamental and applied biological sciences. Psychology
Glycoproteins
Glycoproteins - chemistry
HNK‐1
Molecular Sequence Data
Myelin
Myelin Proteins - chemistry
Myelin Sheath - chemistry
oligosaccharides
Oligosaccharides - chemistry
PASII/PMP22
Peptide Fragments - chemistry
Peripheral Nerves - chemistry
Proteins
Spectrometry, Mass, Fast Atom Bombardment
Sulfated glycoprotein
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Summary: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 Po 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 1H‐NMR studies and comparing the derivatives of PA‐OPPE1 and PA‐oligosaccharides of γ‐globulin on HPLC. The structure, SO4‐3GlcAβ1‐3Galβ1‐4GlcNAcβ1‐2Manα1‐6(GlcNAcβ1‐4) (GlcNAcβ1‐2Manα1‐3)Manβ1‐4GlcNAcβ1‐4(Fucα1‐6)GlcNAc‐PA, was identical to the pyridylaminated form of the major oligosaccharide D8 of bovine Po previously reported.
ISSN:0022-3042
1471-4159
DOI:10.1046/j.1471-4159.2000.0750853.x