Modular interior loading and exterior decoration of a virus-like particle.
Modular interior loading and exterior decoration of a virus-like particle.
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模块化的内部负荷和类似病毒的颗粒的外部装饰。
DOI：
10.1039/c7nr03018e
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发表时间：
2017-07-27
期刊：
影响因子：
6.7
通讯作者：
Douglas T
中科院分区：
文献类型：
--
作者：
Sharma J;Uchida M;Miettinen HM;Douglas T
来源于噬菌体P22的病毒样颗粒（VLP）为货物分子的包封和多价呈递提供了一个有趣且可延展的平台。P22 VLP中货物的包装通常通过遗传激活的定向体内包封来实现。然而，这种方法不允许控制封装货物的包装密度和组成。在这里，我们采用了体外组装方法来控制P22中的货物包装。在体外组装过程中，通过密切调节货物融合支架蛋白和野生型支架蛋白的化学计量比来控制包装。在“一锅组装反应”中，将外壳蛋白亚基与不同比例的野生型支架蛋白和货物融合支架蛋白一起孵育，这导致两种组分包封在共组装的衣壳中。这些实验表明，输入化学计量可用于实现VLP内多种货物的受控包装。P22的多孔性质允许野生型支架蛋白从组装的衣壳逃逸和重新进入，但由于货物的大小，融合至蛋白质-货物的支架蛋白不能穿过衣壳壳。这使得我们能够通过选择性地从共组装的衣壳释放野生型支架蛋白来控制和改变堆积密度。我们已经证明了这些概念在P22系统中使用包封的链霉亲和素蛋白，并已显示其高度选择性的相互作用与生物素或生物素衍生物。此外，该系统可用于包封与生物素偶联的小分子，或展示大蛋白，其不能进入衣壳，因此当连接到柔性生物素化接头时，仍可用于衣壳外部的多价展示。因此，我们已经开发了具有受控蛋白质货物组成和堆积密度的P22系统，小分子和大分子都可以在衣壳的内部或外部以高拷贝数连接到该系统。
Virus-like particles (VLPs) derived from the bacteriophage P22 offer an interesting and malleable platform for encapsulation and multivalent presentation of cargo molecules. The packaging of cargo in P22 VLP is typically achieved through genetically enabled directed in vivo encapsulation. However, this approach does not allow control over the packing density and composition of the encapsulated cargos. Here, we have adopted an in vitro assembly approach to gain control over cargo packaging in P22. The packaging was controlled by closely regulating the stoichiometric ratio of cargo-fused-scaffold protein and wild-type scaffold protein during the in vitro assembly. In a “one-pot assembly reaction” coat protein subunits were incubated with varied ratios of wild-type scaffold protein and cargo-fused-scaffold protein, which resulted in the encapsulation of both components in a co-assembled capsid. These experiments demonstrate that an input stoichiometry can be used to achieve controlled packaging of multiple cargos within the VLP. The porous nature of P22 allows the escape and re-entry of wild-type scaffold protein from the assembled capsid but scaffold protein fused to a protein-cargo cannot traverse the capsid shell due to the size of the cargo. This has allowed us to control and alter the packing density by selectively releasing wild-type scaffold protein from the co-assembled capsids. We have demonstrated these concepts in the P22 system using an encapsulated streptavidin protein and have shown its highly selective interaction with biotin or biotin derivatives. Additionally, this system can be used to encapsulate small molecules coupled to biotin, or display large proteins, that cannot enter the capsid and thus remain available for the multivalent display on the exterior of the capsid when attached to a flexible biotinylated linker. Thus, we have developed a P22 system with controlled protein cargo composition and packing density, to which both small and large molecules can be attached at high copy number on the interior or exterior of the capsid.
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影响因子：
17.1
作者：
Glasgow, Jeff E.;Capehart, Stacy L.;Francis, Matthew B.;Tullman-Ercek, Danielle
通讯作者：
Tullman-Ercek, Danielle
影响因子：
17.1
作者：
Ashley CE;Carnes EC;Phillips GK;Durfee PN;Buley MD;Lino CA;Padilla DP;Phillips B;Carter MB;Willman CL;Brinker CJ;Caldeira Jdo C;Chackerian B;Wharton W;Peabody DS
通讯作者：
Peabody DS
影响因子：
16.6
作者：
Elani, Yuval;Law, Robert V.;Ces, Oscar
通讯作者：
Ces, Oscar
用于估计酸性肽的理论预测分子量和 SDS PAGE 显示分子量之间差异的方程
影响因子：
4.6
作者：
Guan Y;Zhu Q;Huang D;Zhao S;Jan Lo L;Peng J
通讯作者：
Peng J
影响因子：
3.7
作者：
GREENE, B;KING, J
通讯作者：
KING, J