Acosta-Rivero Nelson, Rodriguez Armando, Musacchio Alexis, Falcón Viviana, Suarez Viana M, Martinez Gillian, Guerra Ivis, Paz-Lago Dalila, Morera Yanelys, de la Rosa María C, Morales-Grillo Juan, Dueñas-Carrera Santiago
Hepatitis C Department, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, C.P. 10600, Cuba.
Biochem Biophys Res Commun. 2004 Dec 3;325(1):68-74. doi: 10.1016/j.bbrc.2004.10.012.
Different variants of hepatitis C virus core protein (HCcAg) have proved to self-assemble in vitro into virus-like particles (VLPs). However, difficulties in obtaining purified mature HCcAg have limited these studies. In this study, a high degree of monomeric HCcAg purification was accomplished using chromatographic procedures under denaturing conditions. Size exclusion chromatography and sucrose density gradient centrifugation of renatured HCcAg (in the absence of structured RNA) under reducing conditions suggested that it assembled into empty capsids. The electron microscopy analysis of renatured HCcAg showed the presence of spherical VLPs with irregular shapes and an average diameter of 35nm. Data indicated that HCcAg monomers assembled in vitro into VLPs in the absence of structured RNA, suggesting that recombinant HCcAg used in this work contains all the information necessary for the assembly process. However, they also suggest that some cellular factors might be required for the proper in vitro assembly of capsids.
丙型肝炎病毒核心蛋白（HCcAg）的不同变体已被证明在体外能自组装成病毒样颗粒（VLPs）。然而，获取纯化的成熟HCcAg存在困难，限制了这些研究。在本研究中，通过变性条件下的色谱程序实现了高度纯化的单体HCcAg。在还原条件下对复性的HCcAg（无结构化RNA）进行尺寸排阻色谱和蔗糖密度梯度离心分析表明，它组装成了空衣壳。对复性的HCcAg进行电子显微镜分析显示存在形状不规则、平均直径为35nm的球形VLPs。数据表明，在无结构化RNA的情况下，HCcAg单体在体外组装成VLPs，这表明本研究中使用的重组HCcAg包含组装过程所需的所有信息。然而，这也表明衣壳在体外的正确组装可能需要一些细胞因子。
