ROLE OF SUGAR CHAINS IN THE INVITRO ACTIVITY OF RECOMBINANT HUMAN INTERLEUKIN-5
ROLE OF SUGAR CHAINS IN THE INVITRO ACTIVITY OF RECOMBINANT HUMAN INTERLEUKIN-5
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DOI：
10.1111/j.1432-1033.1993.tb17624.x
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发表时间：
1993-02-01
期刊：
影响因子：
--
通讯作者：
KOBATA, A
中科院分区：
文献类型：
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作者：
KODAMA, S;TSUJIMOTO, M;KOBATA, A
采用高效阴离子交换柱色谱法，在高pH条件下，对中国仓鼠卵巢细胞产生的重组人白细胞介素5（rhIL-5）的O-糖链结构进行了分析。通过对内切-α-N-乙酰氨基半乳糖苷酶的敏感性和对唾液酸键的测定，确定了rhIL-5的核心序列为Galbeta 1->3-GalNAc。为了研究糖链在rhIL-5体外活性中的作用，用各种糖苷酶对其进行了消化。去除N-连接的糖链导致活性增加2.8倍，去-O-糖基化的rhIL-5显示出比完整rhIL-5高10倍的活性，这表明O-连接的糖链的存在比N-连接的糖链更有效地抑制活性。去N-糖基化rhIL-5在70 ℃孵育30 min后活性下降，而完整和去O-糖基化rhIL-5几乎没有活性损失，这表明N-连接的糖链有助于分子的热稳定性。
The structures of 0-linked sugar chains of recombinant human interleukin 5 (rhIL-5) produced by Chinese hamster ovary cells were determined employing high-performance anion-exchange column chromatography under high pH conditions. The core sequence was elucidated to be Galbeta1-->3-GalNAc by its susceptibility to endo-alpha-N-acetylgalactosaminidase and sialic acid linkages were determined using sialidases of different specificities.To investigate the role of sugar chains in the in-vitro activity of rhIL-5, it was digested with various glycosidases. While removal of N-linked sugar chains resulted in 2.8-fold increase of the activity, de-0-glycosylated rhIL-5 showed 10-fold higher activity than intact rhIL-5, suggesting that the presence of 0-linked sugar chains suppresses the activity more effectively than that of N-linked chains. While incubation of de-N-glycosylated rhIL-5 at 70-degrees-C for 30 min decreased the activity, intact and de-0-glycosylated rhIL-5 lost little activity, suggesting that N-linked sugar chains contributed to the thermostability of the molecule.