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J. Biochem, 1988, Vol. 103, No. 6 944-949
© 1988 Japanese Biochemical Society

research-article

Structures of Sugar Chains of Ricin D

Yoshinobu Kimura, Faculty of Agriculture*,1, Sumihiro Hase**, Yuji Kobayashi***, Yoshimasa Kyogoku***, Tokuji Ikenaka** and Gunki Funatsu*,2

*Laboratory of Biochemistry, Kyushu University Higashi-ku, Fukuoka, Fukuoka 812
**Department of Chemistry, Osaka University College of Science Toyonaka, Osaka 560
***Institute for Protein Research, Osaka University Suita, Osaka 565

2To whom correspondence should be addressed.

The toxic lectin, ricin D, contains mannose, fucose, xylose, and N-acetylglucosamine as sugar components. Sugar chains are linked to Asn-10 of the A-chain, and to Asn-95 and Asn-135 of the B-chain (Funatsu, G. et al. (1978) Agric. Biol. Chem. 42, 501-503; Araki, T.& Funatsu, G. (1985) FEBS Lett. 191, 121-124). Asparagine-linked sugar chains of each glycopeptide from ricin D were liberated by hydrazinolysis followed by N-acetylation. The reducing end residues of the sugar chains were coupled with 2-aminopyridine and the pyridylamino (PA-) derivatives obtained were purified by gel-filtration and reversed-phase HPLC. Eight main PA-sugar chains were obtained from three glycopeptides and the structures of these sugar chains were determined by component analysis, stepwise exoglycosidase digestions, partial acetolysis, and 500 MHZ 1H-NMR spectroscopy. The results show that oligomannose type sugar chains (Man6-7GlcNAc2) are linked to Asn-95; Man5-7 GlcNAc2 and M4X (structure, see below) to Asn-135 of the B-chain, and M3FX and M3X to Asn-10 of the A-chain.

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1Present address: Laboratory of Biochemistry, Faculty of Agriculture, Okayama University, Okayama, Okayama 700.


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