A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl beta-D-lactoside derivatives.
4-Methylumbelliferyl 6'-O-benzyl-beta-D-lactoside (6'Bn-MU-Lac) and some related compounds were synthesized via 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, the K(m) value is 0.232 mM at pH 5.
Wang
L X
LX
Department of Biology, Johns Hopkins University, Baltimore, MD 21218-2685, USA.
Pavlova
N V
NV
Li
S C
SC
Li
Y T
YT
Lee
Y C
YC
D000818:Animals; D002855:Chromatography, Thin Layer; D006026:Glycoside Hydrolases / Q000032:analysis* / Q000378:metabolism*; D006027:Glycosides / Q000138:chemical synthesis* / Q000378:metabolism; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D007865:Leeches / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D013050:Spectrometry, Fluorescence / Q000379:methods; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
C102546:4-methylumbelliferyl lactoside; D006027:Glycosides; D007202:Indicators and Reagents; D006923:Hymecromone; D006026:Glycoside Hydrolases; C051111:ceramide glycanase