Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against beta 1-->2 xylose-containing N-glycans.
From the seeds of Ginkgo biloba, a glycoprotein, which is a major component that reacts with an antiserum against beta 1-->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-NAc2)-V-T-V-A-F, which corresponded to the N-terminal amino acid sequence.
Kimura
Y
Y
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Japan. yosh8mar@ccews2.cc.okayama-u.ac.jp
Harada
T
T
Matsuo
S
S
Yonekura
M
M
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D020441:Ginkgo biloba / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D007106:Immune Sera; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010946:Plants, Medicinal; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology*; D011492:Protein Hydrolysates / Q000032:analysis; D012639:Seeds / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014994:Xylose / Q000737:chemistry*
D006023:Glycoproteins; D007106:Immune Sera; D011134:Polysaccharides; D011492:Protein Hydrolysates; D014994:Xylose