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目的：研究人可溶性ＩＬ６Ｒ（ｈｓＩＬ６Ｒ）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ｓＩＬ６Ｒ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Ｗｅｓｔｅｒｎｂｌｏｔ检测证实转染细胞上清中的表达产物，并利用ＢｉｎｄｉｎｇａｓａｙＥＬＩＳＡ和生物学功能分析法检测表达产物与ＩＬ６的结合能力以及它们所介导的ＩＬ６信号转导功能的差异。结果：突变体Ｈ２８０Ｉ比天然ｓＩＬ６Ｒ具有更高的ＩＬ６结合能力，但拮抗ＩＬ６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ｓＩＬ６Ｒ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。
Objective: To study the spatial structure-activity relationship of human soluble IL6R (hsIL6R), providing a theoretical basis for the design of small molecule antagonists. Methods: First of all, PCR amplification of natural human sIL 6R gene fragment, and then the 280th His residue was mutated to Ile. The natural gene and the mutant gene were transfected into COS7 cells respectively, and the expression products in the supernatant of the transfected cells were confirmed by ELISA and Western blotting. The binding ability of the expressed product and IL-6 was detected by Bindingasay ELISA and biological functional analysis Mediated IL 6 signal transduction function difference. Results: The mutant H280I had higher IL-6 binding capacity than the native sIL6R, but antagonized the signal transduction function of IL6 in two effector cell lines. Conclusion: The 280th His residue has an important influence on the normal biological function of sIL6R, which is a good candidate for the antagonist design.