Chemoenzymatic synthesis of neoglycoproteins using transglycosylation with endo-beta-N-acetylglucosaminidase A.
Chemoenzymatic synthesis of neoglycoproteins using transglycosylation with endo-beta-N-acetylglucosaminidase A.
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使用内切 β-N-乙酰氨基葡萄糖苷酶 A 进行转糖基化，化学酶法合成新糖蛋白。
DOI：
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发表时间：
2001
期刊：
影响因子：
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通讯作者：
K. Takegawa
中科院分区：
文献类型：
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作者：
K. Fujita;K. Takegawa
描述了一种新的化学酶法合成含高甘露糖型寡糖的新糖蛋白。利用内切-β-N-乙酰氨基葡萄糖苷酶A（Endo-A）的转糖基化活性，将对异硫氰酸基苯基-β-d-吡喃葡萄糖苷（Glc-ITC）转移到高甘露糖型寡糖的还原末端。合成了一种新的寡糖Man（6）GlcNAc-Glc-ITC，作为蛋白质部分的赖氨酰和N-末端残基的偶联试剂。将新糖缀合物与几种非糖基化蛋白如核糖核酸酶A、溶菌酶和α-乳白蛋白偶联。这些蛋白质的每个分子中含有一到四种高甘露糖型寡糖。该方法对于合成具有同质高甘露糖型低聚糖的新糖蛋白非常有用。
A novel chemoenzymatic approach to synthesize neoglycoproteins containing high-mannose-type oligosaccharides is described. p-Isothiocyanatophenyl-beta-d-glucopyranoside (Glc-ITC) was transferred to the reducing end of the high-mannose-type oligosaccharides using a transglycosylation activity of endo-beta-N-acetylglucosaminidase A (Endo-A). A novel oligosaccharide, Man(6)GlcNAc-Glc-ITC, was synthesized as a coupling reagent for lysyl and N-terminal residues of the protein moiety. The neoglycoconjugate was coupled with several nonglycosylated proteins such as ribonuclease A, lysozyme, and alpha-lactalbumin. Between one and four high-mannose-type oligosaccharides were incorporated per molecule of these proteins. This method should be very useful for the synthesis of neoglycoproteins with homogeneous high-mannose-type oligosaccharides.