Structural Analysis of Free N-Glycans Occurring in Soybean Seedlings and Dry Seeds
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概要
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The structures of unconjugated or free N-glycans in stems of soybean seedlings and dry seeds have been identified. The free N-glycans were extracted from the stems of seedlings or defatted dry seeds. After desalting by two kinds of ion-exchange chromatography and a gel filtration, the free N-glycans were coupled with 2-aminopyridine. The resulting fluorescence-labeled (PA-) N-glycans were purified by gel filtration, Con A affinity chromatography, reverse-phase HPLC, and size-fractionation HPLC. The structures of the PA-sugar chains purified were analyzed by the combination of twodimensional sugar chain mapping, jack bean α-mannosidase digestion, α-1,2-mannosidase digestions, partial acetolysis, and ESI-MS/MS. The free N-glycan structures found showed that two categories of free N-glycans occur in the stems of soybean seedlings. One is a high-mannose type structure having one GlcNAc residue at the reducing end (Man_<9-5>GlcNAc_1,93%), that would be derived by endo-GM (Kimura, Y. et al., Biochim. Biophys. Acta, 1381,27-36 (1998)). The other small component is a xylose-containing type one having two GlcNAc residues at the reducing end (Man_3Xyl_1 GlcNAc_2,7%), which would be derived by PNGase-GM (Kimura, Y. and Ohno, A., Biosci. Biotechnol. Biochem., 62,412-418 (1998)). The detailed structural analysis of free glycans showed that high-mannose type free N-glycans (Man_<9-5>GlcNAc_1) in the soybean seedlings have a common core structural unit ; Manα1-6 (Man1-3) Manα1-6 (Manα1-3) Manβ1-4GlcNAc. Comparing the amount of free N-glycans in the seedling stems and dry seeds, the amount in the stems of seedlings was much higher than that in the dry seeds ; approximately 700 pmol per one stem, 8 pmol in one dry seed. This fact suggested that free N-glycans in soybean seedlings could be produced by two kinds of N-glycan releasing enzymes during germination or seedlingdevelopment.
- 社団法人日本農芸化学会の論文
- 2000-09-23
著者
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Kimura Yoshinobu
Department Of Agricultural Science Faculty Of Agriculture Okayama University
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KITAHARA Erika
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
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Kitahara Erika
Department Of Bioresources Chemistry Faculty Of Agriculture Okayama University
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