从蛋黄中制备天冬酰胺连接的单葡萄糖基化高甘露糖型寡糖。
Preparation of asparagine-linked monoglucosylated high-mannose-type oligosaccharide from egg yolk.
作者信息
Wang Ning, Seko Akira, Takeda Yoichi, Ito Yukishige
机构信息
Japan Science and Technology Agency (JST), ERATO, Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Synthetic Cellular Chemistry Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Japan Science and Technology Agency (JST), ERATO, Ito Glycotrilogy Project, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
出版信息
Carbohydr Res. 2015 Jun 26;411:37-41. doi: 10.1016/j.carres.2015.04.011. Epub 2015 Apr 26.
Monoglucosylated high-mannose-type glycan (Glc1Man9GlcNAc2: G1M9) is well-known as a key glycoform in the glycoprotein folding process, which is specifically recognized by lectin chaperones calnexin (CNX) and calreticulin (CRT) in the endoplasmic reticulum (ER). In this work, we developed an efficient method for the preparation of G1M9-Asn. The G1M9-Asn was obtained from the IgY-rich fraction derived from hen egg yolk by the digestion with pronase. The α-amino group of asparagine in G1M9-Asn was protected with the 9-fluorenylmethyloxycarbonyl (Fmoc) group and the labeled glycans were subsequently purified using high performance liquid chromatography (HPLC). This method will provide useful substrates for analysis of the glycoprotein folding cycle in the ER.
单糖基化高甘露糖型聚糖（Glc1Man9GlcNAc2：G1M9）作为糖蛋白折叠过程中的关键糖型而广为人知，它在内质网（ER）中被凝集素伴侣钙联蛋白（CNX）和钙网蛋白（CRT）特异性识别。在这项工作中，我们开发了一种高效制备G1M9-Asn的方法。通过用链霉蛋白酶消化从鸡蛋黄中获得的富含IgY的部分来得到G1M9-Asn。G1M9-Asn中天冬酰胺的α-氨基用9-芴甲氧羰基（Fmoc）基团保护，随后使用高效液相色谱（HPLC）纯化标记的聚糖。该方法将为内质网中糖蛋白折叠循环的分析提供有用的底物。
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