Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 957-960 |
| Number of pages | 4 |
| Journal | Glycobiology |
| Volume | 9 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - Sep 1 1999 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
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Requirement for a different hydrophobic moiety and reliable chromogenic substrate for endo-type glycosylceramidases. / Miura, Yoshiaki; Arai, Tsutomu; Ohtake, Atsuko; Ito, Makoto; Yamamoto, Kenji; Yamagata, Tatsuya.
In: Glycobiology, Vol. 9, No. 9, 01.09.1999, p. 957-960.Research output: Contribution to journal › Article
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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
PY - 1999/9/1
Y1 - 1999/9/1
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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UR - http://www.scopus.com/inward/citedby.url?scp=0032831458&partnerID=8YFLogxK
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 -