SYNTHESIS OF A DI-O-METHYLATED PENTASACCHARIDE FOR USE IN THE ASSAY OF N-ACETYLGLUCOSAMINYLTRANSFERASE-III ACTIVITY

被引：6
作者：
KHAN, SH [1 ]
COMPSTON, CA [1 ]
PALCIC, MM [1 ]
HINDSGAUL, O [1 ]
机构：
[1] UNIV ALBERTA,DEPT CHEM,EDMONTON T6G 2G2,AB,CANADA
来源：
CARBOHYDRATE RESEARCH | 1994年 / 262卷 / 02期
基金：
英国医学研究理事会; 加拿大自然科学与工程研究理事会;
关键词：
OLIGOSACCHARIDE; PENTASACCHARIDE; N-ACETYLGLYCOSYLAMINYLTRANSFERASE III; BLOCK SYNTHESIS; SUBSTRATE;
D O I：
10.1016/0008-6215(94)84185-3
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The biantennary oligosaccharide analogue beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->3)-[beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->6)]beta-D-Manp-O(CH2)(8)COOMe (3) is a potential substrate for N-acetylglucosaminyltransferases (GlcNAcTs) III-V which are present in mammalian cells. The di-O-methylated analogue of 3, beta-D-GlcpNAc-(1-->2)-[4-O-methyl-alpha-D-Manp]-(1-->3)-[beta-D-GlcpNAc-(1-->2)-[6-O-methyl-alpha-D-Manp]-(1-->6)]-beta-D-Manp-O-(CH2)(8)COOMe (5), was prepared by a block synthesis approach involving sequential addition of two O-methylated disaccharide donors to a protected central beta-D-Man residue. The OH groups acted on by GlcNAcT-IV and -V are protected from glycosylation in 5 since they are present as their methyl ethers. Pentasaccharide 5 was found to be an excellent substrate for GlcNAcT-III (EC 2.4.1.144) from rat kidney with K-m = 0.15 mM. The product formed by incubation of 5 with a rat kidney extract, in the presence of UDP-GlcNAc, was isolated, structurally characterized by NMR spectroscopy and confirmed to be the expected di-O-methyl hexasaccharide where a beta-D-GlcpNAc residue had been added to OH-4 of the central beta-D-Manp unit.
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收藏
页码：283 / 295
页数：13
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