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人可溶性IL |
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宋伦 任蕴芳 王建安 沈倍奋
摘 要 目的：研究人可溶性IL-6R(hsIL-6R)的空间构效关系，为小分子拮抗剂设计提供理论依据。方法：首先利用PCR技术扩增天然人sIL-6R基因片段，然后将第280位His残基突变成Ile。天然基因和突变体基因分别转染COS7细胞，经ELISA和Western blot检测证实转染细胞上清中的表达产物，并利用Binding assay ELISA和生物学功能分析法检测表达产物与IL-6的结合能力以及它们所介导的IL-6信号转导功能的差异。结果：突变体H280I比天然sIL-6R具有更高的IL-6结合能力，但拮抗IL-6在两种效应细胞系上的信号传递功能。结论：第280位His残基对sIL-6R发挥正常生物学功能具有重要影响，是符合拮抗剂设计要求的一个很好的候选位点。
关键词 人可溶性IL-6R(hsIL-6R) 突变体 生物学活性
中国图书分类号 R392.11
Expression and biological activity analysis of human soluble IL-6R and its mutant
SONG Lun，REN Yun-Fang，WANG Jian-An et al.
Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing 100850
Abstract Objective:To study the structure-function relationship of human soluble IL-6 receptor(hsIL-6R) and lay a foundation to design the antagonists.Methods:The sIL-6R gene was obtained by PCR and then histine 280 was mutanted into isolucine with site-directed mutagenesis system.The expression plasmids of the two genes were introduced into COS7 cells and then the expressed products were confirmed by ELISA and Western blot.IL-6 binding abilities were measured by binding assay ELISA.The biological function analysis of the expressed products on the IL-6 responsive cells indicated their different abilities of signal transduction.Results:The mutant H280I had higher IL-6 binding ability than that of wtsIL-6R,but showed antagonistic function on IL-6 signal transdution.Conclusion:H280 played an important role in the function of sIL-6R and might be used as a target site for the antagonist design.
Key words Human soluble IL-6R Mutant Biological activity
IL-6是一种多功能的细胞因子，它的多功能性是由其受体介导的。IL-6R由α、β两条链组成，二者都具有膜结合性和可溶性受体两种形式，α链为配基特异性受体(一般称IL-6R)，β链是IL-6类型的细胞因子的公用转导子，也称GP130［1］，α链具有同IL-6结合并进一步与GP130偶联的能力，它的这种生物学功能是由其胞外区细胞因子结合功能域(也称CBD区)执行的［2］。目前，对CBD区的结构和功能的研究国外已取得了一定进展，并分别确定了该区域中与IL-6结合以及与GP130结合有关的一些氨基酸残基，其中第280位的His残基是影响sIL-6R与GP13[1] [2] 下一页
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