Structural analysis of N-linked carbohydrate chains of funnel web spider (Agelenopsis aperta) venom peptide isomerase.
The structure of the N-linked carbohydrate chains of peptide isomerase from the venom of the funnel web spider (Agelenopsis aperta) has been analyzed. Carbohydrates were released from peptide isomerase by hydrazinolysis and reductively aminated with 2-aminopyridine. The fluorescent derivatives were purified by phenol/chloroform extraction, followed by size-exclusion HPLC. The structure of the purified pyridylamino (PA-) carbohydrate chains were analyzed by a combination of two-dimensional HPLC mapping, sugar composition analysis, sequential exoglycosidase digestions, and mass spectrometry. The peptide isomerase contains six kinds of N-linked carbohydrate chains of truncated high-mannose type, with a fucose alpha 1-6 linked to the reducing N-acetylglucosamine in approximately 80% of them.
Tsukuba Research Laboratories, Eisai Co. Ltd., Ibaraki, Japan. y1-shikata@eisai.co.jp
Y
Y
Shikata
H
H
Ohe
N
N
Mano
M
M
Kuwada
N
N
Asakawa
D000591:Amino Acid Isomerases / Q000737:chemistry*; D000631:Aminopyridines; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D008969:Molecular Sequence Data; D011073:Polyamines / Q000032:analysis*; D013111:Spider Venoms / Q000032:analysis*
D000631:Aminopyridines; C071920:FTX, spider toxin; D011073:Polyamines; D013111:Spider Venoms; D000591:Amino Acid Isomerases; C032439:alpha-aminopyridine