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目的筛选和鉴定RD细胞能与肠道病毒71型(EV71)VP1蛋白结合的受体蛋白,为进一步深入研究EV71病毒受体特性和功能提供实验依据。方法提取RD细胞膜蛋白,双向凝胶电泳分离RD细胞膜蛋白后,通过Far-western印迹实验筛选能与EV71病毒VP1蛋白相互作用的RD细胞膜蛋白,同时设立阴性对照,利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS),对特异性结合的蛋白点进行鉴定。结果通过Far-western印迹检测,从RD细胞筛选到4个能与EV71结合的蛋白点;质谱分析仅鉴定出一个膜蛋白,即stomatin-like protein2(SLP-2)。结论至少有4个蛋白可以与EV71的VP1蛋白相互作用,其中RD细胞膜上的SLP-2蛋白能够与EV71病毒的VP1特异性结合,可能是EV71病毒候选受体或受体辅助分子之一,其在EV71病毒感染RD细胞过程中的作用有待进一步研究。
Objective To screen and identify the receptor proteins that RD cells can bind to VP1 protein of enterovirus 71 (EV71), providing experimental basis for further study on the characteristics and functions of EV71 virus receptor. METHODS: RD cell membrane protein was extracted and two-dimensional gel electrophoresis was used to separate the RD cell membrane protein. After screening by Far-western blotting, RD cell membrane protein interacting with VP71 protein of EV71 virus was screened. At the same time, a negative control was set up. Matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS) to identify specifically bound protein spots. Results Four protein spots binding to EV71 were screened from RD cells by Far-western blotting. Only one membrane protein, stomatin-like protein2 (SLP-2), was identified by mass spectrometry. Conclusions At least four proteins can interact with the VP1 protein of EV71, in which the SLP-2 protein on RD cell membrane can specifically bind VP1 of EV71 virus, which may be one of EV71 virus candidate receptors or receptor accessory molecules. The role of EV71 virus infection in RD cells needs further study.