RD细胞肠道病毒71型受体的筛选  

Identification of enterovirus 71 putative receptor from RD cells

作　　者：胡小云[1] 王长兵[1] 朱冰[1] 钟家禹[1] 邝璐[1] 

机构地区：[1]广州医学院附属广州市妇女儿童医疗中心中心实验室,广东广州510120

出　　处：《中华医院感染学杂志》2012年第10期2022-2025,共4页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金项目(30972630)

摘　　要：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 identify the enterovirus 71（EV71） VP1 binding proteins from EV71-sensitive RD cells and provide experimental evidence for the following virus receptor research.METHODS The cell membrane from permissive RD cells was isolated by using sequential protein extraction,then was solubilized followed by using SDS-PAGE and two dimensional（2D） gel electrophoresis to separate the membrane proteins and applied a modified western blot（Far-western blotting） to show the VP1 binding proteins,the identification of binding proteins of specificity was performed by matrix-assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS）.RESULTS Far-western blotting test showed that four cellular proteins of RD cells were capable of binding with VP1,and only one membrane protein which was stomatin-like protein 2（SLP-2） was identified by mass spectrometry.CONCLUSION At least 4 proteins interact with the VP1 of the EV71,in which SLP-2 can bind specifically to EV71,and may be a putative receptor for EV71 and its role in RD cells infected with EV71 remains to be studied further.

关 键 词：肠道病毒71型 双向电泳 Far-western印迹 飞行时间质谱 SLP-2 

分 类 号：R373.2[医药卫生—病原生物学]