TY - JOUR
T1 - Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells
T2 - Predominant occurrence of complex type N-glycans
AU - Maeda, Megumi
AU - Kimura, Yoshinobu
N1 - Funding Information:
* This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Basic Research C, no. 17580300), and the Okayama University COE program ‘‘Establishment of Plant Health Science.’’ y To whom correspondence should be addressed. Fax: +81-86-251-8296; E-mail: yosh8mar@cc.okayama-u.ac.jp Abbreviations: PA-, pyridylamino; RP-HPLC, reverse-phase HPLC; SF-HPLC, size-fractionation HPLC; ESI-MS, electrospray ionization mass spectrometry; MS/MS, tandem mass; endo-β-GlcNAc-ase, endo-β-N-acetylglucosaminidase; Hex, hexose; Pen, pentose; deoxyHex, deoxy hexose; HexNAc, N-acetyl hexosamine; M3FX, Manα1-6(Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; GNM3FX, GlcNAcβ1-2Manα1-6(Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; GN2M3FX, GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; G2F2GN2M3FX, Galβ1-3(Fucα1-4)GlcNAcβ1-2Manα1-6(Galβ1-3(Fucα1-4)GlcNAcβ1-2Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; M5A, Manα1-6(Manα1-3)Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA; M6B, Manα1-6(Manα1-3)Manα1-6(Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA; M7A, Manα1-2Manα1-6(Manα1-3)Manα1-6(Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA; M8A, Manα1-2Manα1-6(Manα1-3)Manα1-6(Manα1-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA; M8C, Manα1-6(Manα1-2Manα1-3)Manα1-6(Manα1-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA; M9A, Manα1-2Manα1-6(Manα1-2Manα1-3)Manα1-6(Manα1-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-PA
PY - 2006
Y1 - 2006
N2 - Although it has been found that plant endo-β-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-β-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man 9-5GlcNAc1) occur at concentration of several micromolar (μM). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.
AB - Although it has been found that plant endo-β-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-β-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man 9-5GlcNAc1) occur at concentration of several micromolar (μM). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.
KW - N-glycan metabolism
KW - N-glycan structures
KW - Oriza sativa
KW - Rice glycoprotein
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U2 - 10.1271/bbb.50637
DO - 10.1271/bbb.50637
M3 - Article
C2 - 16794314
AN - SCOPUS:33745483793
VL - 70
SP - 1356
EP - 1363
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 6
ER -