从人乳中纯化的β-D-半乳糖基转移酶的复合型天冬酰胺连接糖链的结构鉴定。
Structure identification of the complex-type, asparagine-linked sugar chains of beta-D-galactosyl-transferase purified from human milk.
作者信息
Endo T, Amano J, Berger E G, Kobata A
出版信息
Carbohydr Res. 1986 Aug 1;150:241-63. doi: 10.1016/0008-6215(86)80020-9.
The asparagine-linked sugar chains of human milk galactosyltransferase were quantitatively released as oligosaccharides from the polypeptide backbone by hydrazinolysis. They were converted into radioactive oligosaccharides by sodium borotritiate reduction after N-acetylation, and fractionated by paper electrophoresis and by Bio-Gel P-4 column chromatography after sialidase treatment. Structural studies of each oligosaccharides by sequential exoglycosidase digestion and methylation analysis indicated that the galactosyltransferase contains bi, tri-, and probably tetra-antennary, complex-type oligosaccharides having alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-beta-D-Manp -(1----4)-beta-D- GlcpNac-(1----4)-alpha-L-[Fucp-(1----6)]-D-GlcNAc as their common core. Variation is produced by the different locations and numbers of the five different outer chains: beta-D-Galp-(1----4)-D-GlcNAc, alpha-L-Fucp-(1----3)-[beta-D-Galp-(1----4)]-D-GlcNAc, alpha-NeuAc-(2----6)-beta-D-Galp-(1----4)-D-GlcNAc, alpha-L-Fucp-(1----3)-[beta-D-Galp-(1----4)]-beta-D-GlcpNAc-(1---- 3)-beta- D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-D-GlcNAc, and alpha-NeuAc-(2----6)-beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----3)-beta-D - Galp-(1----4)-[alpha-L-Fucp-(1----3)]-beta-D-GlcNAc.
人乳半乳糖基转移酶的天冬酰胺连接糖链通过肼解从多肽主链上定量释放为寡糖。经N - 乙酰化后，通过硼氢化三钠还原将它们转化为放射性寡糖，并在唾液酸酶处理后通过纸电泳和Bio - Gel P - 4柱色谱进行分级分离。通过顺序外切糖苷酶消化和甲基化分析对每种寡糖进行结构研究表明，半乳糖基转移酶含有具有α - D - 甘露糖基 - (1→3)-[α - D - 甘露糖基 - (1→6)]-β - D - 甘露糖基 - (1→4)-β - D - N - 乙酰葡糖胺 - (1→4)-α - L - [岩藻糖基 - (1→6)]-D - 葡糖胺作为其共同核心的双天线、三天线以及可能的四天线复合型寡糖。五种不同外链在位置和数量上的差异产生了变化：β - D - 半乳糖基 - (1→4)-D - N - 乙酰葡糖胺、α - L - 岩藻糖基 - (1→3)-[β - D - 半乳糖基 - (1→4)]-D - N - 乙酰葡糖胺、α - N - 乙酰神经氨酸 - (2→6)-β - D - 半乳糖基 - (1→4)-D - N - 乙酰葡糖胺、α - L - 岩藻糖基 - (1→3)-[β - D - 半乳糖基 - (1→4)]-β - D - N - 乙酰葡糖胺基 - (1→3)-β - D - 半乳糖基 - (1→4)-[α - L - 岩藻糖基 - (1→3)]-D - N - 乙酰葡糖胺以及α - N - 乙酰神经氨酸 - (2→6)-β - D - 半乳糖基 - (1→4)-β - D - N - 乙酰葡糖胺基 - (1→3)-β - D - 半乳糖基 - (1→4)-[α - L - 岩藻糖基 - (1→3)]-β - D - N - 乙酰葡糖胺。
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