Abstract:
RNA bacteriophage MS2-derived virus-like particles (VLPs) have been widely used in biomedical research as model systems to study virus assembly, structure-function relationships, vaccine development, and drug delivery. Considering the diverse utility of these VLPs, a systemic engineering approach has been utilized to generate smaller particles with optimal serum stability and tissue penetrance. Additionally, it is crucial to demonstrate the overall stability of these mini MS2 VLPs, ensuring cargo protection until they reach their target cell/organ. However, no detailed analysis of the thermal stability and heat-induced disassembly of MS2 VLPs has yet been attempted. In this work, we investigated the thermal stability of both wild-type (WT) MS2 VLP and its \"mini\" variant containing S37P mutation (mini MS2 VLP). The mini MS2 VLP exhibits a higher capsid melting temperature (Tm) when compared to its WT MS2 VLP counterpart, possibly attributed to its smaller interdimer angle. Our study presents that the thermal unfolding of MS2 VLPs follows a sequential process involving particle destabilization, nucleic acid exposure/melting, and disassembly of VLP. This observation underscores the disruption of cooperative intersubunit interactions and protein-nucleic acid interactions, shedding light on the mechanism of heat-induced VLP disassembly.
摘要:
RNA噬菌体MS2衍生的病毒样颗粒(VLP)已广泛用于生物医学研究,作为研究病毒组装的模型系统,结构-功能关系,疫苗开发,和药物输送。考虑到这些VLP的不同效用,一种系统工程方法已被用来产生具有最佳血清稳定性和组织渗透度的较小颗粒。此外,证明这些迷你MS2VLP的整体稳定性至关重要,确保货物保护,直到他们到达目标细胞/器官。然而,尚未尝试对MS2VLP的热稳定性和热诱导分解进行详细分析。在这项工作中,我们研究了野生型(WT)MS2VLP及其含有S37P突变的“迷你”变体(迷你MS2VLP)的热稳定性。与WTMS2VLP对应物相比,迷你MS2VLP表现出较高的衣壳熔化温度(Tm),可能归因于其较小的二聚体间角度。我们的研究表明,MS2VLP的热展开遵循涉及颗粒不稳定的连续过程,核酸暴露/熔解,和VLP的拆卸。这一观察结果强调了合作亚基间相互作用和蛋白质-核酸相互作用的破坏,光对热诱导VLP拆卸机理的研究。