Ohta M, Hamako J, Yamamoto S, Hatta H, Kim M, Yamamoto T, Oka S, Mizuochi T, Matsuura F
Department of Biotechnology, Faculty of Engineering, Fukuyama University, Hiroshima, Japan.
Glycoconj J. 1991 Oct;8(5):400-13. doi: 10.1007/BF00731292.
Asparagine-linked oligosaccharides present on hen egg-yolk immunoglobulin, termed IgY, were liberated from the protein by hydrazinolysis. After N-acetylation, the oligosaccharides were labelled with a UV-absorbing compound, p-aminobenzoic acid ethyl ester (ABEE). The ABEE-derivatized oligosaccharides were fractionated by anion exchange, normal phase and reversed phase HPLC, and their structures were determined by a combination of sugar composition analysis, methylation analysis, negative ion FAB-MS, 500 MHz 1H-NMR and sequential exoglycosidase digestions. IgY contained monoglucosylated oligomannose type oligosaccharides with structures of Glc alpha 1-3Man7-9-GlcNAc-GlcNAc, oligomannose type oligosaccharides with the size range of Man5-9GlcNAc-GlcNAc, and biantennary complex type oligosaccharides with core region structure of Man alpha 1-6(+/- GlcNAc beta 1-4)(Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4(+/- Fuc alpha 1-6)GlcNAc. The glucosylated oligosaccharides, Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2, have not previously been reported in mature glycoproteins from any source.
存在于鸡卵黄免疫球蛋白（称为IgY）上的天冬酰胺连接的寡糖通过肼解从蛋白质中释放出来。N-乙酰化后，寡糖用一种紫外线吸收化合物对氨基苯甲酸乙酯（ABEE）进行标记。ABEE衍生的寡糖通过阴离子交换、正相和反相高效液相色谱进行分离，其结构通过糖组成分析、甲基化分析、负离子快原子轰击质谱、500兆赫1H-核磁共振和顺序外切糖苷酶消化相结合的方法确定。IgY含有结构为Glcα1-3Man7-9-GlcNAc-GlcNAc的单葡萄糖基化寡甘露糖型寡糖、大小范围为Man5-9GlcNAc-GlcNAc的寡甘露糖型寡糖以及核心区域结构为Manα1-6(+/-GlcNAcβ1-4)(Manα1-3)Manβ1-4GlcNAcβ1-4(+/-Fucα1-6)GlcNAc的双触角复合型寡糖。葡萄糖基化寡糖Glc1Man8GlcNAc2和Glc1Man7GlcNAc2以前在任何来源的成熟糖蛋白中都未被报道过。
