TY - JOUR
T1 - A novel disaccharide substrate having 1,2-oxazoline moiety for detection of transglycosylating activity of endoglycosidases
AU - Fujita, Masaya
AU - Shoda, Shin ichiro
AU - Haneda, Katsuji
AU - Inazu, Toshiyuki
AU - Takegawa, Kaoru
AU - Yamamoto, Kenji
N1 - Funding Information:
This work was partially supported by a Grant-in-Aid for Scientific Research on Priority Area (No. 10134204) from the Ministry of Education, Culture, Sports, Science and Technology, Japan. S.S. gratefully acknowledges financial support from the Tokuyama Science Foundation.
PY - 2001/9/3
Y1 - 2001/9/3
N2 - A disaccharide substrate of Manβ1-4GlcNAc-oxazoline 2 was designed and synthesized as a novel probe for detection of the transglycosylating activity of endoglycosidases. A regio- and stereoselective transglycosylation reaction of 2 to GlcNAcβ1-O-pNP or Dns-Asn(GlcNAc)-OH catalyzed by endo-β-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) and endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (Endo-A) has been demonstrated for the first time, resulting in the core trisaccharide derivative Manβ1-4GlcNAcβ1-4GlcNAcβ1-O-pNP 8 (or -(Dns)Asn-OH). Interestingly, the transglycosylation proceeds irreversibly; the resulting trisaccharide 8 was not hydrolyzed by Endo-M and Endo-A. Based on these results, a new mechanism including an oxazolinium ion intermediate has been proposed for the endoglycosidase-catalyzed hydrolysis or transglycosylation.
AB - A disaccharide substrate of Manβ1-4GlcNAc-oxazoline 2 was designed and synthesized as a novel probe for detection of the transglycosylating activity of endoglycosidases. A regio- and stereoselective transglycosylation reaction of 2 to GlcNAcβ1-O-pNP or Dns-Asn(GlcNAc)-OH catalyzed by endo-β-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) and endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (Endo-A) has been demonstrated for the first time, resulting in the core trisaccharide derivative Manβ1-4GlcNAcβ1-4GlcNAcβ1-O-pNP 8 (or -(Dns)Asn-OH). Interestingly, the transglycosylation proceeds irreversibly; the resulting trisaccharide 8 was not hydrolyzed by Endo-M and Endo-A. Based on these results, a new mechanism including an oxazolinium ion intermediate has been proposed for the endoglycosidase-catalyzed hydrolysis or transglycosylation.
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U2 - 10.1016/S0304-4165(01)00164-7
DO - 10.1016/S0304-4165(01)00164-7
M3 - Article
C2 - 11514092
AN - SCOPUS:0035801838
SN - 0304-4165
VL - 1528
SP - 9
EP - 14
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 1
ER -