衣壳
单体
凝胶电泳
聚丙烯酰胺凝胶电泳
低聚物
染色
二硫键
电泳
牛乳头状瘤病毒
生物
分子生物学
重组DNA
类病毒颗粒
考马斯亮蓝
生物物理学
生物化学
化学
酶
聚合物
高分子化学
基因
基因组
有机化学
遗传学
作者
Qinjian Zhao,Helen H. Guo,Yang Wang,Michael W. Washabaugh,Robert D. Sitrin
标识
DOI:10.1016/j.jviromet.2005.03.015
摘要
The recombinant major capsid protein (L1) of human papillomavirus (HPV) can self-assemble into virus-like particles (VLPs) with 360 L1 molecules per VLP. These tightly associated L1 oligomers in the assembled VLPs were disrupted in a pH-, denaturant-, time-, and temperature-dependent fashion. With non-reducing Laemmli-type SDS-PAGE, primarily the monomeric L1 protein ( approximately 55 kDa) is observed when analyzing VLP preparations. When the pH was lowered to pH 7.0 in NuPAGE system and the gel temperature during electrophoresis was maintained at a lower temperature ( approximately 7 degrees C), a ladder of protein bands in approximately 55 kDa increments were detected above the monomeric p55 band. These discrete bands visualized as a ladder are likely the disulfide-linked L1 oligomers. In addition to the gel running conditions, an increase in pH, temperature, or SDS concentration during sample treatment was also shown to significantly reduce the amount of detectable oligomers, further corroborating the labile nature of these oligomers. Altogether, the results also implicate the redox-responsive nature of the HPV capsid comprising of >95% L1 protein. Molecular basis of the facile disulfide bond inter-change is discussed. This electrophoretic technique for trapping the disulfide-linked oligomers may be employed to detect the oligomeric status of other protein aggregates or assembled particles.
科研通智能强力驱动
Strongly Powered by AbleSci AI