Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase

Yohei Ishibashi, Yusuke Nagamatsu, Sandra Meyer, Akihiro Imamura, Hideharu Ishida, Makoto Kiso, Nozomu Okino, Rudolf Geyer, Makoto Ito

研究成果: ジャーナルへの寄稿記事

8 引用 (Scopus)

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
19
発行部数7
DOI
出版物ステータス出版済み - 6 8 2009

Fingerprint

Glycosphingolipids
Glycolipids
Labeling
Oligosaccharides
Pentanols
Fungi
Parasites
Taenia
Rhizopus
Sphingolipids
Mollusca
Ceramides
Metabolic Networks and Pathways
Hydrolysis
High Pressure Liquid Chromatography
Detectors
Substrates

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

Ishibashi, Y, Nagamatsu, Y, Meyer, S, Imamura, A, Ishida, H, Kiso, M, Okino, N, Geyer, R & Ito, M 2009, 'Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase', Glycobiology, 巻. 19, 番号 7, pp. 797-807. https://doi.org/10.1093/glycob/cwp051
Ishibashi, Yohei ; Nagamatsu, Yusuke ; Meyer, Sandra ; Imamura, Akihiro ; Ishida, Hideharu ; Kiso, Makoto ; Okino, Nozomu ; Geyer, Rudolf ; Ito, Makoto. / Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase. :: Glycobiology. 2009 ; 巻 19, 番号 7. pp. 797-807.
@article{e9c72153a3b44e1b9a92153a4dba47b0,
title = "Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase",
abstract = "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.",
author = "Yohei Ishibashi and Yusuke Nagamatsu and Sandra Meyer and Akihiro Imamura and Hideharu Ishida and Makoto Kiso and Nozomu Okino and Rudolf Geyer and Makoto Ito",
year = "2009",
month = "6",
day = "8",
doi = "10.1093/glycob/cwp051",
language = "English",
volume = "19",
pages = "797--807",
journal = "Glycobiology",
issn = "0959-6658",
publisher = "Oxford University Press",
number = "7",

}

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 -