N-glycan structures of murine hippocampus serine protease, neuropsin, produced in Trichoplusia ni cells.
N-glycans of neuropsin (serine protease in the murine hippocampus) expressed in Trichoplusia ni cells were released from the glycopeptides by digestion with glycoamidase A (from sweet almond), and the reducing ends of the oligosaccharides were reductively aminated with 2-aminopyridine. The derivatized N-glycans were separated and structurally identified by a two dimensional high-performance liquid chromatography (HPLC) mapping technique on two kinds of HPLC columns. Fourteen different major N-glycan structures were identified, of which 6 were high-mannose type (9.1%), and the remaining 8 were paucimannosidic type. The presence of insect specific N-glycan structures containing both alpha1,3- and alpha1,6- di-fucosylated innermost N-acetylglucosamine residue (23.3%), as below, was also confirmed by 600 MHz 1H-NMR spectroscopy.
Takahashi
N
N
GlycoLab, Mitsukan Group Co., Ltd., Aichi, Japan. ntakahas@mb.infoweb.ne.jp
Tsukamoto
Y
Y
Shiosaka
S
S
Kishi
T
T
Hakoshima
T
T
Arata
Y
Y
Yamaguchi
Y
Y
Kato
K
K
Shimada
I
I
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002240:Carbohydrate Sequence; D006624:Hippocampus / Q000201:enzymology*; D007610:Kallikreins; D007915:Lepidoptera / Q000166:cytology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics
D011134:Polysaccharides; D011994:Recombinant Proteins; C094799:KLK8 protein, human; D007610:Kallikreins; D012697:Serine Endopeptidases