Abstract
From the seeds of Ginkgo biloba, a glycoprotein, which is a major component that reacts with an antiserum against β1→2 xylose-containing N-glycans, has been purified and characterized. The N-terminal amino acid sequence of the purified glycoprotein was H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-L-L-F-G-Q-. The molecular mass was estimated to be 17 kDa and 16 kDa by SDS-PAGE under reducing conditions, however, the molecular mass of this glycoprotein in the native state was 30,762 by MALDI-TOF MS, suggesting that this glycoprotein consists of two subunits; one is glycosylated and the other is not. The structure of N-glycan linked to this glycoprotein (designated 30 kDa GBGP) was identified as Man3Fuc1Xyl1GlcNAc2, which is the predominant N-glycan linked to the storage glycoproteins in the same seeds (Kimura, Y et al. (1998) Biosci. Biotechnol. Biochem. 62, 253-261). From the peptic digest of the carboxymethylated glycosylated subunit, one glycopeptide was purified by RP-HPLC and the amino acid sequence was identified as H-K-A-N-N(Man3Fuc1Xyl1Glc-NAc 2)-V-T-V-A-F, which corresponded to the N-terminal amino acid sequence.
| Original language | English |
|---|---|
| Pages (from-to) | 463-467 |
| Number of pages | 5 |
| Journal | Bioscience, Biotechnology and Biochemistry |
| Volume | 63 |
| Issue number | 3 |
| Publication status | Published - Mar 1999 |
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Keywords
- Antigenic N-glycan
- Antiserum against plant N-glycan
- Ginkgo biloba
- Immunoblotting
- Plant glycoprotein
ASJC Scopus subject areas
- Food Science
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biotechnology
- Chemistry (miscellaneous)
- Applied Microbiology and Biotechnology
- Bioengineering
Cite this
Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against β1→2 xylose-containing N-glycans. / Kimura, Yoshinobu; Harada, Takuo; Matsuo, Sayuri; Yonekura, Masami.
In: Bioscience, Biotechnology and Biochemistry, Vol. 63, No. 3, 03.1999, p. 463-467.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against β1→2 xylose-containing N-glycans
AU - Kimura, Yoshinobu
AU - Harada, Takuo
AU - Matsuo, Sayuri
AU - Yonekura, Masami
PY - 1999/3
Y1 - 1999/3
N2 - From the seeds of Ginkgo biloba, a glycoprotein, which is a major component that reacts with an antiserum against β1→2 xylose-containing N-glycans, has been purified and characterized. The N-terminal amino acid sequence of the purified glycoprotein was H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-L-L-F-G-Q-. The molecular mass was estimated to be 17 kDa and 16 kDa by SDS-PAGE under reducing conditions, however, the molecular mass of this glycoprotein in the native state was 30,762 by MALDI-TOF MS, suggesting that this glycoprotein consists of two subunits; one is glycosylated and the other is not. The structure of N-glycan linked to this glycoprotein (designated 30 kDa GBGP) was identified as Man3Fuc1Xyl1GlcNAc2, which is the predominant N-glycan linked to the storage glycoproteins in the same seeds (Kimura, Y et al. (1998) Biosci. Biotechnol. Biochem. 62, 253-261). From the peptic digest of the carboxymethylated glycosylated subunit, one glycopeptide was purified by RP-HPLC and the amino acid sequence was identified as H-K-A-N-N(Man3Fuc1Xyl1Glc-NAc 2)-V-T-V-A-F, which corresponded to the N-terminal amino acid sequence.
AB - From the seeds of Ginkgo biloba, a glycoprotein, which is a major component that reacts with an antiserum against β1→2 xylose-containing N-glycans, has been purified and characterized. The N-terminal amino acid sequence of the purified glycoprotein was H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-L-L-F-G-Q-. The molecular mass was estimated to be 17 kDa and 16 kDa by SDS-PAGE under reducing conditions, however, the molecular mass of this glycoprotein in the native state was 30,762 by MALDI-TOF MS, suggesting that this glycoprotein consists of two subunits; one is glycosylated and the other is not. The structure of N-glycan linked to this glycoprotein (designated 30 kDa GBGP) was identified as Man3Fuc1Xyl1GlcNAc2, which is the predominant N-glycan linked to the storage glycoproteins in the same seeds (Kimura, Y et al. (1998) Biosci. Biotechnol. Biochem. 62, 253-261). From the peptic digest of the carboxymethylated glycosylated subunit, one glycopeptide was purified by RP-HPLC and the amino acid sequence was identified as H-K-A-N-N(Man3Fuc1Xyl1Glc-NAc 2)-V-T-V-A-F, which corresponded to the N-terminal amino acid sequence.
KW - Antigenic N-glycan
KW - Antiserum against plant N-glycan
KW - Ginkgo biloba
KW - Immunoblotting
KW - Plant glycoprotein
UR - http://www.scopus.com/inward/record.url?scp=0033089308&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033089308&partnerID=8YFLogxK
M3 - Article
C2 - 10227132
AN - SCOPUS:0033089308
VL - 63
SP - 463
EP - 467
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 3
ER -