Asn-linked sugar chain structures of recombinant human thrombopoietin produced in Chinese hamster ovary cells
Asn-linked sugar chain structures of recombinant human thrombopoietin produced in Chinese hamster ovary cells
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DOI：
10.1023/a:1007159409961
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发表时间：
1999-11-01
影响因子：
3
通讯作者：
Takeuchi, M
中科院分区：
文献类型：
--
作者：
Inoue, N;Watanabe, T;Takeuchi, M
人促红细胞生成素（TPO）是一种与人促红细胞生成素（EPO）部分同源的N-和o -糖基化糖蛋白激素，调节巨核细胞和血小板的数量。利用2-氨基苯甲酰胺标记法、序列糖苷酶切法和基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF/MS）对重组人TPO的糖链结构进行定量分析。我们发现双、三和四天线复合物型糖链具有一个或两个n -乙酰乳胺重复序列，这是CHO细胞中产生的重组人EPO所共有的。另一方面，具有1个或2个n -乙酰乳胺重复序列的三天线糖链是重组人TPO所特有的，它们的分支结构分布也不同。这些结果表明，除了糖基转移酶亚群外，近端蛋白质结构还决定了asn链的分支结构。
Human thrombopoietin (TPO) that regulates the numbers of megakaryocytes and platelets is a heavily N- and O-glycosylated glycoprotein hormone with partial homology to human erythropoietin (EPO). We prepared recombinant human TPO produced in Chinese hamster ovary (CHO) cells and analyzed the sugar chain structures quantitatively using 2-aminobenzamide labeling, sequential glycosidase digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS).We found bi-, tri- and tetraantennary complex-type sugar chains with one or two N-acetyllactosamine repeats, which are common to recombinant human EPO produced in CHO cells. On the other hand, there were triantennary sugar chains with one or two N-acetyllactosamine repeats that were specific to the recombinant human TPO, and their distributions of branch structures were also different. These results suggested that proximal protein structure should determine the branch structure of Asn-linked sugar chains in addition to the glycosyltransferases subset.