Collection of α1-acid glycoprotein molecular species by capillary electrophoresis and the analysis of their molecular masses and carbohydrate chains -: Basic studies on the analysis of glycoprotein glycoforms
Collection of α1-acid glycoprotein molecular species by capillary electrophoresis and the analysis of their molecular masses and carbohydrate chains -: Basic studies on the analysis of glycoprotein glycoforms
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DOI：
10.1016/s0021-9673(02)00353-9
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发表时间：
2002-06-07
影响因子：
4.1
通讯作者：
Kakehi, K
中科院分区：
文献类型：
--
作者：
Sei, K;Nakano, M;Kakehi, K
在含有0.5%（w/v）羟丙基甲基纤维素的20 mM醋酸盐缓冲液（pH 4.2）中，使用表面改性毛细管，通过毛细管电泳将高度异质性糖蛋白（α（1）-酸性糖蛋白）拆分为糖型。我们收集的馏分含有每种糖型作为近纯态的毛细管电泳，并检查这些糖型的分子量的基质辅助激光解吸飞行时间质谱。用N-糖苷酶F释放糖链，然后用8-氨基芘-1，3，6-三磺酸盐荧光标记糖链，分析糖型的分离，发现糖型的分离主要是由于多触角糖链的存在。我们建议，本技术是有用的分析蛋白质的翻译后修饰的碳水化合物链。(C)2002 Elsevier Science B. V.保留所有权利。
A highly heterogeneous glycoprotein, alpha(1)-acid glycoprotein, was resolved into their glycoforms by capillary electrophoresis using a surface-modified capillary in 20 mM acetate buffer (pH 4.2) containing 0.5% (w/v) hydroxypropylmethylcellulose. We collected the fractions containing each glycoform as nearly pure state by capillary electrophoresis, and examined the molecular masses of these glycoforms by matrix assisted laser desorption time-of-flight mass spectrometry. We also analyzed carbohydrate chains after releasing them with N-glycosidase F followed by fluorescent labeling with 8-aminopyrene-1,3,6-trisulfonate, We found that the separation of glycoforms was mostly due to the presence of multiantennary carbohydrate chains. We propose that the present technique is useful for the analysis of post translational modification of proteins with carbohydrate chains. (C) 2002 Elsevier Science B.V. All rights reserved.