抄録
Although 6-gala series glycosphingolipids possessing R-Gal (α/ β) 1-6Galβ1-1′Cer have been found in some mollusks, pathogenic parasites, and fungi, their physiological functions and metabolic pathway are not fully understood. We described a novel method of detecting 6-gala series glyco- sphingolipids utilizing the specificity of endogalactosylceramidase (EGALC), which is capable of hydrolyzing 6-gala series glycosphingolipids to produce intact oligosaccharides and ceramides. EGALC catalyzes not only hydrolysis but also a transglycosylation reaction. In the latter reaction, EGALC transfers oligosaccharides from the glycosphingolipids to acceptors such as fluorescent 1-alkanols. Based on the transglycosylation reaction of EGALC, a specific, easy, fast, sensitive, and reproducible method of detecting 6-gala series glycosphingolipids was developed using NBD-pentanol as an acceptor. The fluorescent products, NBD-pentanol-conjugated 6-gala oligosaccharides, were separated and detected by TLC or HPLC with a fluorescent detector. Moreover, it was revealed that as well as glycosphingolipids, a glycoglycerolipid, digalactosyldiacylglycerol, was utilized by EGALC as a donor substrate. This method was successfully applied to detect 6-gala series glycosphingolipids in a fungus, Rhizopus oryzae, and a parasite, Taenia crassiceps. The method would be useful for studying glycosphingolipids and galactosyl glycerolipids which share the Gal (α/β) 1-6Gal structure.
| 元の言語 | 英語 |
|---|---|
| ページ（範囲） | 797-807 |
| ページ数 | 11 |
| ジャーナル | Glycobiology |
| 巻 | 19 |
| 発行部数 | 7 |
| DOI | |
| 出版物ステータス | 出版済み - 6 8 2009 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
これを引用
Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase. / Ishibashi, Yohei; Nagamatsu, Yusuke; Meyer, Sandra; Imamura, Akihiro; Ishida, Hideharu; Kiso, Makoto; Okino, Nozomu; Geyer, Rudolf; Ito, Makoto.
：: Glycobiology, 巻 19, 番号 7, 08.06.2009, p. 797-807.研究成果: ジャーナルへの寄稿 › 記事
}
TY - JOUR
T1 - Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase
AU - Ishibashi, Yohei
AU - Nagamatsu, Yusuke
AU - Meyer, Sandra
AU - Imamura, Akihiro
AU - Ishida, Hideharu
AU - Kiso, Makoto
AU - Okino, Nozomu
AU - Geyer, Rudolf
AU - Ito, Makoto
PY - 2009/6/8
Y1 - 2009/6/8
N2 - Although 6-gala series glycosphingolipids possessing R-Gal (α/ β) 1-6Galβ1-1′Cer have been found in some mollusks, pathogenic parasites, and fungi, their physiological functions and metabolic pathway are not fully understood. We described a novel method of detecting 6-gala series glyco- sphingolipids utilizing the specificity of endogalactosylceramidase (EGALC), which is capable of hydrolyzing 6-gala series glycosphingolipids to produce intact oligosaccharides and ceramides. EGALC catalyzes not only hydrolysis but also a transglycosylation reaction. In the latter reaction, EGALC transfers oligosaccharides from the glycosphingolipids to acceptors such as fluorescent 1-alkanols. Based on the transglycosylation reaction of EGALC, a specific, easy, fast, sensitive, and reproducible method of detecting 6-gala series glycosphingolipids was developed using NBD-pentanol as an acceptor. The fluorescent products, NBD-pentanol-conjugated 6-gala oligosaccharides, were separated and detected by TLC or HPLC with a fluorescent detector. Moreover, it was revealed that as well as glycosphingolipids, a glycoglycerolipid, digalactosyldiacylglycerol, was utilized by EGALC as a donor substrate. This method was successfully applied to detect 6-gala series glycosphingolipids in a fungus, Rhizopus oryzae, and a parasite, Taenia crassiceps. The method would be useful for studying glycosphingolipids and galactosyl glycerolipids which share the Gal (α/β) 1-6Gal structure.
AB - Although 6-gala series glycosphingolipids possessing R-Gal (α/ β) 1-6Galβ1-1′Cer have been found in some mollusks, pathogenic parasites, and fungi, their physiological functions and metabolic pathway are not fully understood. We described a novel method of detecting 6-gala series glyco- sphingolipids utilizing the specificity of endogalactosylceramidase (EGALC), which is capable of hydrolyzing 6-gala series glycosphingolipids to produce intact oligosaccharides and ceramides. EGALC catalyzes not only hydrolysis but also a transglycosylation reaction. In the latter reaction, EGALC transfers oligosaccharides from the glycosphingolipids to acceptors such as fluorescent 1-alkanols. Based on the transglycosylation reaction of EGALC, a specific, easy, fast, sensitive, and reproducible method of detecting 6-gala series glycosphingolipids was developed using NBD-pentanol as an acceptor. The fluorescent products, NBD-pentanol-conjugated 6-gala oligosaccharides, were separated and detected by TLC or HPLC with a fluorescent detector. Moreover, it was revealed that as well as glycosphingolipids, a glycoglycerolipid, digalactosyldiacylglycerol, was utilized by EGALC as a donor substrate. This method was successfully applied to detect 6-gala series glycosphingolipids in a fungus, Rhizopus oryzae, and a parasite, Taenia crassiceps. The method would be useful for studying glycosphingolipids and galactosyl glycerolipids which share the Gal (α/β) 1-6Gal structure.
UR - http://www.scopus.com/inward/record.url?scp=66249112827&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=66249112827&partnerID=8YFLogxK
U2 - 10.1093/glycob/cwp051
DO - 10.1093/glycob/cwp051
M3 - Article
C2 - 19389917
AN - SCOPUS:66249112827
VL - 19
SP - 797
EP - 807
JO - Glycobiology
JF - Glycobiology
SN - 0959-6658
IS - 7
ER -