Multifunctional Enzyme Packaging and Catalysis in the Qβ Protein Nanoparticle,Biomacromolecules

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Multifunctional Enzyme Packaging and Catalysis in the Qβ Protein Nanoparticle
Biomacromolecules ( IF 5.5 ) Pub Date : 2018-08-30 00:00:00 , DOI: 10.1021/acs.biomac.8b00885
Jason D. Fiedler ₁ , Maxwell R. Fishman ₁ , Steven D. Brown ₁ , Jolene Lau ₁ , M. G. Finn _{1,

2}

Affiliation

The simultaneous expression in Escherichia coli cells of the Qβ virus-like particle (VLP) capsid protein and protein “cargo” tagged with a positively charged Rev peptide sequence leads to the spontaneous self-assembly of VLPs with multiple copies of the cargo inside. We report the packaging of four new enzymes with potential applications in medicine and chemical manufacturing. The captured enzymes are active while inside the nanoparticle shell and are protected from environmental conditions that lead to free-enzyme destruction. We also describe genetic modifications to the packaging scheme that shed light on the self-assembly mechanism of this system and allow indirect control over the internal packaging density of cargo. The technology was extended to create, via self-assembly, VLPs that simultaneously display protein ligands on the exterior and contain enzymes within. Inverse relationships were observed between the size of both the packaged and externally displayed protein or domains and nanoparticle yield. These results provide a general method for the rapid creation of robust protein nanoparticles with desired catalytic and targeting functionalities.

中文翻译：

Qβ蛋白纳米颗粒中的多功能酶包装和催化

在大肠杆菌中同时表达Qβ病毒样颗粒（VLP）衣壳蛋白和带有正电荷Rev肽序列标记的蛋白“货物”的细胞会导致VLP的自发自组装，并在其中带有多个拷贝的货物。我们报告了四种新酶的包装，这些新酶在医药和化学制造中具有潜在的应用前景。所捕获的酶在纳米颗粒壳内部时是有活性的，并受到保护，免受导致自由酶破坏的环境条件的影响。我们还描述了对包装方案的基因改造，从而揭示了该系统的自组装机制，并允许间接控制货物的内部包装密度。该技术已扩展为通过自组装创建VLP，这些VLP同时在外部显示蛋白质配体并在其中包含酶。在包装的和外部展示的蛋白质或结构域的大小与纳米颗粒产量之间观察到反比关系。这些结果提供了一种快速创建具有所需催化和靶向功能的坚固蛋白质纳米颗粒的通用方法。

更新日期：2018-08-30

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