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A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl β-d-lactoside derivatives

  • Papers Dedicated To Dr Sen-Itiroh Hakomori
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Abstract

4-Methylumbelliferyl 6′-O-benzyl-β-d-lactoside (6′Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6′Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK m value is 0.232 mM at pH 5.

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Authors and Affiliations

  1. Department of Biology, The Johns Hopkins University, 21218-2685, Baltimore, MD, USA

    Lai-Xi Wang & Yuan C. Lee

  2. Department of Biochemistry, Tulane University Medical Center, 70112-2699, New Orleans, LO, USA

    Nadejda V. Pavlova, Su-Chen Li & Yu-Teh Li

Authors
  1. Lai-Xi Wang
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  2. Nadejda V. Pavlova
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  3. Su-Chen Li
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  4. Yu-Teh Li
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  5. Yuan C. Lee
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Additional information

Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl β-d-lactoside; b′Bn-Lac, 6′-O-benzyl-lactose; 6′Bn-MU-Lac, 4-methylumbelliferyl 6′-Obenzyl-β-d-lactoside; 4′6′Bd-MU-Lac, 4-methylumbelliferyl 4′,6′-O-benzylidene-β-d-lactoside; MU-Cel, 4-methylumbellifery β-d-cellobioside; 4′6′Bd-MU-Cel, 4-methylumbelliferyl 4′,6′-O-benzylidene-β-d-cellobioside; TLC, thin layer chromatography;1H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures.

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Wang, LX., Pavlova, N.V., Li, SC. et al. A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl β-d-lactoside derivatives. Glycoconjugate J 13, 359–365 (1996). https://doi.org/10.1007/BF00731468

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  • DOI: https://doi.org/10.1007/BF00731468

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