Comparative structural study of N-linked oligosaccharides of urinary and recombinant erythropoietins.
Comparative structural study of N-linked oligosaccharides of urinary and recombinant erythropoietins.
复制标题
尿促红细胞生成素和重组促红细胞生成素的 N 连接寡糖的比较结构研究。
作者：
E. Tsuda;M. Goto;A. Murakami;K. Akai;M. Ueda;G. Kawanishi;N. Takahashi;R. Sasaki;H. Chiba;H. Ishihara
本文分析了从再生障碍性贫血患者尿中纯化的尿促红细胞生成素（u-EPO）的N-连接寡糖的结构，并与用幼仓鼠肾（BHK）细胞制备的重组促红细胞生成素（r-EPO）进行了比较。通过N-寡糖糖肽酶（杏仁）消化从每个EPO蛋白中释放天冬酰胺连接的中性寡糖。将由此获得的寡糖链的还原末端用荧光试剂2-氨基吡啶胺化，并将寡糖的吡啶基氨基衍生物的混合物通过高效液相色谱法（HPLC）在ODS硅胶柱上分离。一步高效液相色谱法可将u-EPO和r-EPO（BHK）分别分离出8种和13种以上的寡糖组分。通过连续外切糖苷酶消化和另一种具有酰胺-硅胶柱的HPLC的组合来分析由此分离的每种寡糖的结构。此外，高分辨率质子核磁共振（1H NMR）光谱和甲基化分析进行的情况下，r-EPO（BHK）。（250字处删节）
The structures of the N-linked oligosaccharides of the urinary erythropoietin (u-EPO) purified from urine of aplastic anemic patients were analyzed and compared with those for recombinant erythropoietin (r-EPO) prepared with baby hamster kidney (BHK) cells. Asparagine-linked neutral oligosaccharides were released from each EPO protein by N-oligosaccharide glycopeptidase (almond) digestion. The reducing ends of the oligosaccharide chains thus obtained were aminated with a fluorescent reagent, 2-aminopyridine, and the mixture of pyridylamino derivatives of the oligosaccharides was separated by high-performance liquid chromatography (HPLC) on an ODS silica column. More than 8 and 13 kinds of oligosaccharide fractions for u-EPO and r-EPO (BHK), respectively, were completely separated by the one-step HPLC procedure. The structure of each oligosaccharide thus isolated was analyzed by a combination of sequential exoglycosidase digestion and another kind of HPLC with an amide-silica column. Furthermore, high-resolution proton nuclear magnetic resonance (1H NMR) spectroscopy and methylation analyses were carried out in the case of r-EPO (BHK).(ABSTRACT TRUNCATED AT 250 WORDS)
DOI：
10.1016/s0021-9258(18)45317-3
发表时间：
1987-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hiroshi Sasakis;B. Bothner;A. Dell;Minoru Fukudall
通讯作者：
Hiroshi Sasakis;B. Bothner;A. Dell;Minoru Fukudall
人促红细胞生成素基因：在稳定转染的哺乳动物细胞中高水平表达和染色体定位。
DOI：
10.1073/pnas.83.17.6465
发表时间：
1986
影响因子：
11.1
作者：
Powell,JS;Berkner,KL;Lebo,RV;Adamson,JW
通讯作者：
Adamson,JW
N-连接牛胎球蛋白糖肽与分离的兔肝细胞的结合：三触角结构中 Gal/GalNAc 肝凝集素区分 Gal beta(1,4)GlcNAc 和 Gal beta(1,3)GlcNAc。
影响因子：
2.9
作者：
Townsend,RR;Hardy,MR;Wong,TC;Lee,YC
通讯作者：
Lee,YC