人肿瘤坏死因子(hTNFα)30位和42位突变体生物活性差异的结构机制

Structural Mechanism Studies of hTNFαMutants in Position30and42Amino Acid

目的探索人肿瘤坏死因子(hTNFα)结构与功能活性的关系。方法利用免疫沉淀试验、mRNA差异显示等方法初步探索了所构建的几个突变体的结构特点和其作用于细胞后所导致的基因反应差异。结果表现不同生物学活性的突变体其结构存在一定的差异,并且所导致的细胞基因反应程度和种类不完全一致。结论hTNFα诱导的细胞凋亡过程是通过一整套的基因反应来实现的,hTNFα突变体导致的不同生物活性,一些是以其结构因素诱导不同程度的凋亡基因反应来实现的,而另一些则是通过程度各异且种类不同的基因反应而实现的。

Objective To study the relationship between the structure and functional activity of hTNF α .Methods Four hTNF α mutants were constructed,different binding structures and gene responses rel - ated with these mutants were studied by the methods of immunoprecipitation and mRNA differential display.Results The specific activities and LD50of the different hTNF α mutants indicated their dif - ferent bioactivities.It was shown that the hTNF α mutants had the relative binding affinities to the wild types.The mRNA differential display assay proved that the hTNF α mutants stimulated different gene responses.Conclusion These results suggest that the specific anti - tumor activities of hTNF α mutants are accomplished by inducing different or same gene response at different quantities after its binding to specific receptor.