Analysis of Fluorogenic Smith Degradation Products of 7-(1,3-Disulfonaphtyl)amino-Disaccharides for Linkage Position Analysis of Carbohydrates.
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概要
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The linkage position of a glycosidic bond to the reducing-end residue of a pyridylamino (PA-) sugar can be determined sensitively by Smith degradation and HPLC [K. Omichi and S. Hase, (1994) J. Biochem. 115, 429-434]. With the aim of enhancing the sensitivity of this method of linkage position analysis to the fmol-level, use of the 7-(1, 3-disulfonaphtyl)amino (DSNA-) group instead of the PA-group as a fluorescent tag was examined. Smith degradation of DSNA-disaccharides with a DSNA-hexose, DSNA-N-acetylglu-cosamine, or DSNA-N-acetylgalactosamine reducing-end residue was carried out. HPLC and FAB-MS of the fluorogenic Smith degradation products showed that the DSNA-group was stable under the Smith degradation reaction conditions, and that the reaction proceeded in a manner similar to that using PA-disaccharides to give the predicted products. Fluorogenic Smith degradation products specific to the glycosidic linkage position were well separated by reversed-phase HPLC, and were easily assignable by comparing the HPLC elution positions with those of standard compounds. The method was successfully applied to analyzing the structure of an N-linked sugar chain.
- 社団法人 日本生化学会の論文
著者
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Omichi Kaoru
Department Of Applied Chemistry Waseda University
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Makino Yasushi
Department Of Chemistry Graduate School Of Science Osaka Prefecture University
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Yonezaki Kayoko
Department Of Environmental Sciences Faculty Of Science Osaka Women's University
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