Characterization of asparagine-linked oligosaccharides on a mouse submandibular mucin.
Characterization of asparagine-linked oligosaccharides on a mouse submandibular mucin.
复制标题
小鼠颌下粘蛋白上天冬酰胺连接的寡糖的表征。
DOI：
10.1093/glycob/5.6.589
复制
发表时间：
1995
期刊：
影响因子：
4.3
通讯作者：
Hong-Le,NH
中科院分区：
文献类型：
--
作者：
Denny,PC;Denny,PA;Hong-Le,NH
用肽-N4-（N-乙酰-β-氨基葡萄糖）天冬酰胺酶F或内切-β-N-乙酰氨基葡萄糖糖苷酶H处理成年雌性小鼠下颌下腺粘蛋白，释放天冬酰胺连接寡糖。内切-β-N-乙酰氨基葡萄糖苷酶H似乎比肽-N4-（N-乙酰-β-氨基葡萄糖基）-天冬酰胺酰胺酶F更有效地从该粘蛋白释放天冬酰胺连接的寡糖。用荧光团8-氨基萘-1，3，6-磺酸定量还原标记后，用聚丙烯酰胺凝胶电泳分离寡糖。通过一组重组外切糖苷酶对单个寡糖进行测序。大约50%的低聚糖是高甘露糖型。这个家族的五个甘露糖成员是最普遍的。第二组低聚糖是非二等分杂交型。未检测到复杂的天冬酰胺连接寡糖。杂种表现出双触角和三触角分枝模式。三触角杂种是最常见的杂种，占所有寡糖的>30%。由于约98%的杂合寡糖唾液酸化，并且全部缺乏二等分N-乙酰葡糖胺，这些寡糖作为一个组在其他糖蛋白中很少观察到。完全唾液酸化的三触角杂种可能是独特的。
The asparagine-linked oligosaccharides from an adult female mouse submandibular gland mucin were released by treatment with peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase F or endo-β-N-acetylglucosaminidase H. Endo-β-N-acetylglucosaminidase H appeared to be more effective at releasing the asparagine-linked oligosaccharides from this mucin than was peptide-N4-(N-acetyl-β-glucosaminyl)-asparagine amidase F. After quantitative reductive labelling with the fluorophore, 8-aminonaphthalene-1, 3, 6-sulphonic acid, the oligosaccharides were separated by polyacrylamide gel electrophoresis and isolated. The individual oligosaccharides were sequenced by a battery of recombinant exoglycosidases. Approximately 50% of the oligosaccharides were of the high-mannose type. The five-mannose member of this family was the most prevalent. The second group of oligosaccharides were of the non-bisected hybrid type. No complex asparagine-linked oligosaccharides were detected. The hybrids exhibited both biantennary and triantennary branching patterns. The triantennary hybrid was the most common hybrid at >30% of all oligosaccharides. With ∼98% of the hybrid oligosaccharides sialylated and all lacking a bisectingN-acetylglucosamine, these oligosaccharides as a group have been only rarely observed in other glycoproteins. The fully sialylated triantennary hybrid may be unique.