Symmetry Controlled, Genetic Presentation of Bioactive Proteins on the P22 Virus-like Particle Using an External Decoration Protein.
Symmetry Controlled, Genetic Presentation of Bioactive Proteins on the P22 Virus-like Particle Using an External Decoration Protein.
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DOI：
10.1021/acsnano.5b03360
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发表时间：
2015-09-22
期刊：
影响因子：
17.1
通讯作者：
Douglas T
中科院分区：
文献类型：
--
作者：
Schwarz B;Madden P;Avera J;Gordon B;Larson K;Miettinen HM;Uchida M;LaFrance B;Basu G;Rynda-Apple A;Douglas T
病毒利用组成蛋白的空间控制作为操纵和逃避宿主免疫系统的手段。同样，对封装或呈现在设计的纳米粒子上的蛋白质进行精确的空间控制有可能仿生地放大或屏蔽生物相互作用。此前，我们已经展示了将多种客体蛋白封装在来自鼠伤寒沙门氏菌噬菌体 P22 的病毒样颗粒 (VLP) 中的能力，包括来自流感等人类病原体的抗原蛋白。扩展了这种强大的封装策略，我们使用来自噬菌体 L 的三聚体装饰蛋白 (Dec) 作为成熟 P22 VLP 上受控外部呈递的手段，它以高亲和力与成熟 P22 VLP 结合。通过与 Dec 蛋白 C 末端的基因融合，鼠 CD40L 的 17 kDa 可溶区或根据“自我标记”CD47 的结合区设计的最小肽独立地呈现在 P22 VLP 衣壳外部。当以 Dec 融合形式呈现时，两种候选药物都保留了功能。通过表面等离振子共振测量，Dec 结构域与 P22 衣壳的结合在设计的构建体中变化最小，这证明了该呈现策略的广泛实用性。 Dec介导的呈现提供了一种强大的、模块化的方法来装饰P22衣壳的暴露外部，以进一步协调对生物系统内内部功能化的VLP的反应。
Viruses use spatial control of constituent proteins as a means of manipulating and evading host immune systems. Similarly, precise spatial control of proteins encapsulated or presented on designed nanoparticles has the potential to biomimetically amplify or shield biological interactions. Previously, we have shown the ability to encapsulate a wide range of guest proteins within the virus-like particle (VLP) from Salmonella typhimurium bacteriophage P22, including antigenic proteins from human pathogens such as influenza. Expanding on this robust encapsulation strategy, we have used the trimeric decoration protein (Dec) from bacteriophage L as a means of controlled exterior presentation on the mature P22 VLP, to which it binds with high affinity. Through genetic fusion to the C-terminus of the Dec protein, either the 17 kDa soluble region of murine CD40L or a minimal peptide designed from the binding region of the “self-marker” CD47 was independently presented on the P22 VLP capsid exterior. Both candidates retained function when presented as a Dec-fusion. Binding of the Dec domain to the P22 capsid was minimally changed across designed constructs, as measured by surface plasmon resonance, demonstrating the broad utility of this presentation strategy. Dec-mediated presentation offers a robust, modular means of decorating the exposed exterior of the P22 capsid in order to further orchestrate responses to internally functionalized VLPs within biological systems.