Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against β1→2 xylose-containing N-glycans

Yoshinobu Kimura, Takuo Harada, Sayuri Matsuo, Masami Yonekura

Research output: Contribution to journalArticle

9 Citations (Scopus)

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 languageEnglish
Pages (from-to)463-467
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume63
Issue number3
Publication statusPublished - Mar 1999

Fingerprint

Ginkgo biloba
Glycoproteins
Xylose
xylose
Chemical properties
Purification
Polysaccharides
antiserum
Immune Sera
glycoproteins
physicochemical properties
polysaccharides
Amino acids
Amino Acid Sequence
amino acid sequences
Molecular mass
Amino Acids
Seed
Seeds
molecular weight

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 journalArticle

@article{db2190bd42c342e1be0c5e97acc9471b,
title = "Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against β1→2 xylose-containing N-glycans",
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.",
keywords = "Antigenic N-glycan, Antiserum against plant N-glycan, Ginkgo biloba, Immunoblotting, Plant glycoprotein",
author = "Yoshinobu Kimura and Takuo Harada and Sayuri Matsuo and Masami Yonekura",
year = "1999",
month = "3",
language = "English",
volume = "63",
pages = "463--467",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "3",

}

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 -