Engineering of vault nanocapsules with enzymatic and fluorescent properties
Engineering of vault nanocapsules with enzymatic and fluorescent properties
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DOI：
10.1073/pnas.0500929102
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发表时间：
2005-03-22
影响因子：
11.1
通讯作者：
Rome, LH
中科院分区：
文献类型：
--
作者：
Kickhoefer, VA;Garcia, Y;Rome, LH
新兴的纳米技术领域面临的核心问题之一是细胞兼容性。纳米粒子已被提出用于诊断和治疗应用，包括药物输送、基因治疗、生物传感器和受控催化。病毒、脂质体、多肽以及合成和天然聚合物已被设计用于这些应用，但显著的限制继续阻碍它们的使用。避免人体的自然免疫系统，缺乏靶向性，以及无法控制包装和释放仍然是障碍。我们探索了一种自然产生的细胞纳米颗粒的使用，这种纳米颗粒被称为拱顶，它的名字来自于它的形态，它的多个拱门让人想起大教堂的天花板。金库是含有13-丙二醛的核糖核蛋白颗粒，其内部空腔大到足以隔离数百种蛋白质。在这里，我们报告了一种在拱顶腔内靶向和隔离生物活性物质的策略。两种蛋白质--荧光素酶和GFP的一个变种--结合了一个针对拱顶的多肽，导致它们被隔离在拱顶腔内。靶蛋白赋予重组金库酶和荧光特性，这两种特性都可以通过发光来检测。改装后的拱顶与活细胞兼容。设计具有设计特性和功能的拱顶粒子的能力代表着朝着开发生物兼容纳米胶囊迈出的重要一步。
One of the central issues facing the emerging field of nanotechnology is cellular compatibility. Nanoparticles have been proposed for diagnostic and therapeutic applications, including drug delivery, gene therapy, biological sensors, and controlled catalysis. Viruses, liposomes, peptides, and synthetic and natural polymers have been engineered for these applications, yet significant limitations continue to prevent their use. Avoidance of the body's natural immune system, lack of targeting specificity, and the inability to control packaging and release are remaining obstacles. We have explored the use of a naturally occurring cellular nanoparticle known as the vault, which is named for its morphology with multiple arches reminiscent of cathedral ceilings. Vaults are 13-MDa ribonucleoprotein particles with an internal cavity large enough to sequester hundreds of proteins. Here, we report a strategy to target and sequester biologically active materials within the vault cavity. Attachment of a vault-targeting peptide to two proteins, luciferase and a variant of GFP, resulted in their sequestration within the vault cavity. The targeted proteins confer enzymatic and fluorescent properties on the recombinant vaults, both of which can be detected by their emission of light. The modified vaults are compatible with living cells. The ability to engineer vault particles with designed properties and functionalities represents an important step toward development of a biocompatible nanocapsule.