TY - JOUR
T1 - Requirement for a different hydrophobic moiety and reliable chromogenic substrate for endo-type glycosylceramidases
AU - Miura, Yoshiaki
AU - Arai, Tsutomu
AU - Ohtake, Atsuko
AU - Ito, Makoto
AU - Yamamoto, Kenji
AU - Yamagata, Tatsuya
N1 - Funding Information:
We thank Dr. Toshinori Sato at Tokyo Institute of Technology for his kind guidance in the synthesis of lactosides, and we are indebted to SNOW BRAND for providing standard LacCer. This work was supported in part by funds from the Mitsubishi Chemical Corporation and Seikagaku Kogyo Corporation, the Fujisawa Foundation, Life Science Foundation of Japan; grants-in-aid for scientific priority areas (7558092 and 09240104) from the Ministry of Education, Science, and Culture of Japan (T.Y.); and a Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists (Y.M.).
PY - 1999/9
Y1 - 1999/9
N2 - A series of synthetic lactosides with aglycones that differed in length and structure were used to determine the substrate specificity of endo-type glycosylceramidases. Endoglycoceramidases (EGCase) from bacteria preferred lactosides with an acylamide structure over simple n-alkyl lactosides. While ceramide glycanase (CGase) from leech did not show preference, N-Acylaminoethyl β-lactosides and n-alkyl lactosides were substrates for both EGCase and CGase, but N-acylaminobutyl β-lactosides, whose acylamide residue differs from that in ceramide, were not hydrolyzed by EGCases. Thus, EGCases, but not CGase, appear to require an N-acyl group at the same position as that of intact glycosphingolipid for substrate recognition. A p-nitrophenyl lactoside derivative possessing an N-acyl chain was degraded by both EGCases and CGase and this chromogenic substrate may be an alternative substrate for endo-type glycosylceramidase activity. K(m) of the chromogenic lactoside for CGase and Rhodococcus EGCase were 28 μM and 2.9 mM, respectively.
AB - A series of synthetic lactosides with aglycones that differed in length and structure were used to determine the substrate specificity of endo-type glycosylceramidases. Endoglycoceramidases (EGCase) from bacteria preferred lactosides with an acylamide structure over simple n-alkyl lactosides. While ceramide glycanase (CGase) from leech did not show preference, N-Acylaminoethyl β-lactosides and n-alkyl lactosides were substrates for both EGCase and CGase, but N-acylaminobutyl β-lactosides, whose acylamide residue differs from that in ceramide, were not hydrolyzed by EGCases. Thus, EGCases, but not CGase, appear to require an N-acyl group at the same position as that of intact glycosphingolipid for substrate recognition. A p-nitrophenyl lactoside derivative possessing an N-acyl chain was degraded by both EGCases and CGase and this chromogenic substrate may be an alternative substrate for endo-type glycosylceramidase activity. K(m) of the chromogenic lactoside for CGase and Rhodococcus EGCase were 28 μM and 2.9 mM, respectively.
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U2 - 10.1093/glycob/9.9.957
DO - 10.1093/glycob/9.9.957
M3 - Article
C2 - 10460837
AN - SCOPUS:0032831458
VL - 9
SP - 957
EP - 960
JO - Glycobiology
JF - Glycobiology
SN - 0959-6658
IS - 9
ER -