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利用圆二色谱(Circular dichroism,CD)和流式细胞术(Flow cytometry,FCM)分析干扰素(Interferon,IFN)-α的构象与其抗病毒活性之间的关系。以重组人IFN-α(rIFN-α2b和rIFN-α2a)为材料,变温CD谱分析表明,在65℃以下,两种IFN-α的二级结构相对稳定,当温度高于65℃以后,两者的二级结构发生不可逆的改变。FCM分析构象变化前后IFN-α的抗病毒活性发现,构象的改变一定程度上减弱了其抗病毒活性。rIFN-α2b和rIFN-α2a具有相似的变化趋势。以上结果表明,IFN-α的构象是影响其抗病毒活性的重要因素之一,同时也验证了CD结合FCM是分析蛋白质药物构象与其细胞水平上生物学活性相关性的较好组合。
The relationship between the conformation of interferon (IFN) -α and its antiviral activity was analyzed by Circular dichroism (CD) and flow cytometry (FCM). The temperature-dependent CD spectra of recombinant human IFN-α (rIFN-α2b and rIFN-α2a) showed that the secondary structure of IFN-α was relatively stable below 65 ℃. When the temperature was above 65 ℃, Secondary structure of people irreversible change. The antiviral activity of IFN-α before and after the conformational changes was analyzed by FCM. It was found that conformational changes reduced its antiviral activity to a certain extent. Similar trends were observed for rIFN-α2b and rIFN-α2a. The above results indicate that the conformation of IFN-α is one of the important factors affecting its antiviral activity, and it is also verified that CD-bound FCM is a good combination of analyzing the correlation between protein conformation and its biological activity at the cellular level.