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目的:获得SARS冠状病毒小信封蛋白E(E蛋白),研究其性质,分析其可能的功能。方法:用PCR方法构建了E蛋白的质粒,在大肠杆菌中表达蛋白,用圆二色谱(CD)测定蛋白质的二级结构,用生物信息学方法预测蛋白质的二级结构和可能的功能,用分子模拟方法模建E蛋白的三维结构。结果:获得了纯度较高的E蛋白,并用CD测定了其二级结构性质,由CD谱推测了二级结构信息与生物信息学预测的基本一致。生物信息学分析表明,与其他冠状病毒相比,SARS冠状病毒的E蛋白一级结构和二级结构非常保守,跨膜区尤其保守。其功能可能与鼠科肝炎冠状病毒和哺乳动物,冠状病毒类似。结论:SARS冠状病毒E蛋白的表达成功,为研究其结构与功能以及研究SARS冠状病毒本身的性能奠定了基础;在膜环境和水溶液中,E蛋白以类似的方式折叠,但在水溶液中,其第一个β-片由于失去与膜表面的氢键作用而变成随机结构。
OBJECTIVE: To obtain SARS coronavirus small envelope protein E (E protein), study its properties and analyze its possible functions. Methods: The plasmid of E protein was constructed by PCR and the protein was expressed in E. coli. The secondary structure of protein was determined by circular dichroism (CD). The secondary structure and possible function of protein were predicted by bioinformatics method. Molecular modeling method for modeling the three-dimensional structure of E protein. Results: The higher purity of E protein was obtained and its secondary structure was determined by CD. The CD spectra suggested that the secondary structure information and bioinformatics prediction were basically consistent. Bioinformatics analysis showed that compared with other coronaviruses, the primary structure and secondary structure of E protein of SARS coronavirus are very conservative and the transmembrane region is particularly conservative. Its function may be similar to murine hepatitis coronavirus and mammalian, coronavirus. Conclusion: The successful expression of SARS-CoV E protein laid the foundation for studying the structure and function of SARS-CoV, and studying the performance of SARS coronavirus itself. In membrane environment and aqueous solution, E protein is folded in a similar way, but in aqueous solution, The first β-sheet becomes a random structure due to the loss of hydrogen bonding with the membrane surface.