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马尔堡病毒的RNA基因组编码7种结构蛋白,其中核蛋白在基因组的复制和核衣壳形成中发挥重要作用,成为十分有吸引力的抗病毒药物潜在靶标.实验利用亲和质谱技术从甘草提取物中筛选马尔堡核蛋白的配体.首先从甘草粗提物中发现了可能与蛋白结合的配体甘草查尔酮A.在此基础上,对甘草中已知组分的单体混合物和甘草查尔酮A单体分别进行亲和质谱分析,验证了粗提物的质谱筛选结果.而后进一步利用SPR实验确认甘草查尔酮A单体对核蛋白靶标的结合作用,进而通过一系列生化实验发现该单体对马尔堡核蛋白热稳定性和构象均能产生功能性影响.实验发展的亲和质谱技术为从中草药提取物中直接筛选新型抗病毒药物分子提供了简便高效的新方法.
The RNA genome of Marburg virus encodes seven structural proteins, of which nucleoprotein plays an important role in genome duplication and nucleocapsid formation and becomes a potential target of antiviral drugs.Experiment using affinity mass spectrometry extracted from licorice Screening of Marburg nuclear protein ligands.First of all from licorice crude extract was found to be the protein binding ligand licochalcone A. On this basis, the licorice known components of the monomer mixture and Licorice chalcone A monomer were analyzed by affinity mass spectrometry to verify the mass spectrometry results of the crude extract.And then further use of SPR experiments confirm licorice chalcone A monomer binding to the nuclear protein target, and then through a series of biochemical The experimental results showed that the monomer exerted a functional influence on the thermal stability and conformation of Marburg nuclear protein.The experimental development of affinity mass spectrometry provided a simple and efficient new method for the direct screening of novel antiviral drug molecules from Chinese herbal extracts.