Acosta-Rivero N, Aguilar J C, Musacchio A, Falcón V, Viña A, de la Rosa M C, Morales J
Division of Vaccines, Physical Chemistry Department, Electron Microscopy Laboratory, Center for Genetic Engineering and Biotechnology, Ave. 31 e/158 y 190, C.P. 10600, C. Havana, Cuba.
Biochem Biophys Res Commun. 2001 Sep 14;287(1):122-5. doi: 10.1006/bbrc.2001.5561.
Little is known about the mechanism of hepatitis C virion assembly. So the capacity of the entire Hepatitis C virus core protein (HCcAg) produced in Pichia pastoris to form particles either in its native soluble state or after detergent treatment of HCcAg associated to cell debris were studied. Size exclusion chromatography suggested that HCcAg assembled into high molecular weight structures. HCcAg was also specifically recognized by a serum from a chronic HCV carrier patient. This antigen migrated with buoyant density values similar to those obtained for native nucleocapsid particles from infected patients when analyzed using sucrose density gradient centrifugation. The analysis by electron microscopy of purified HCcAg showed aggregates resembling virus-like particles (VLPs) with an average diameter of 30 nm. These results indicated that the HCcAg obtained from P. pastoris assembled into VLPs resembling HCV nucleocapsid particles in a mature stage. Such HCcAg aggregates characterized here could be a valuable tool to elucidate the mechanisms of HCV nucleocapsid assembly.
关于丙型肝炎病毒粒子组装机制知之甚少。因此，研究了在毕赤酵母中产生的整个丙型肝炎病毒核心蛋白（HCcAg）在其天然可溶状态下或对与细胞碎片相关的HCcAg进行去污剂处理后形成颗粒的能力。尺寸排阻色谱表明HCcAg组装成高分子量结构。HCcAg也被一名慢性HCV携带者患者的血清特异性识别。当使用蔗糖密度梯度离心分析时，该抗原的浮力密度值与从感染患者获得的天然核衣壳颗粒的浮力密度值相似。对纯化的HCcAg进行电子显微镜分析，显示出类似于病毒样颗粒（VLP）的聚集体，平均直径为30nm。这些结果表明，从毕赤酵母获得的HCcAg组装成了类似于成熟阶段丙型肝炎病毒核衣壳颗粒的VLP。这里表征的这种HCcAg聚集体可能是阐明丙型肝炎病毒核衣壳组装机制的有价值工具。
