The structure of (xylose)2glucose-O-serine 53 found in the first epidermal growth factor-like domain of bovine blood clotting factor IX.
We reported the presence of a new trisaccharide composed of two xylose and reducing terminal glucose residues linked to serine residues of bovine blood clotting factors VII and IX (Hase, S., Kawabata, S., Nishimura, H., Takeya, H., Sueyoshi, T., Miyata, T., Iwanaga, S., Takao, T., Shimonishi, Y., and Ikenaka, T. (1988) J. Biochem. (Tokyo) 104, 867-868). The present paper describes the detailed structural analysis of the trisaccharide. Glycopeptides were prepared from bovine factor IX by digestion with Pronase followed by purification by column chromatography. The trisaccharide was released from the protein by the beta-elimination reaction with hydrazine, and the reducing end of the sugar chain was tagged with 2-aminopyridine. The fluorescent pyridylamino derivative of the trisaccharide was purified by gel filtration and reversed-phase high performance liquid chromatography. The glycopeptides and pyridylamino-trisaccharide thus obtained were subjected to methylation study, 500-MHz 1H nuclear magnetic resonance spectroscopy, and periodate oxidation. Glucose and xylose belong to the D series by high performance liquid chromatography on a chiral column. From the results, the structure of the trisaccharide is proposed as: D-Xyl p alpha 1-3-D-Xyl p alpha 1-3-D-Glcp beta 1-O-Ser-53.
Hase
S
S
Department of Chemistry, Osaka University College of Science, Japan.
Nishimura
H
H
Kawabata
S
S
Iwanaga
S
S
Ikenaka
T
T
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002850:Chromatography, Gel; D004815:Epidermal Growth Factor; D005164:Factor IX; D006023:Glycoproteins / Q000097:blood; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D012694:Serine
D006023:Glycoproteins; D009844:Oligosaccharides; D012694:Serine; D004815:Epidermal Growth Factor; D005164:Factor IX