Protein cages that readily encapsulate active enzymes of interest present useful nanotools for delivery and catalysis, wherein those with programmable disassembly characteristics serve as particularly attractive platforms. Here, a general guest packaging system based on an artificial protein cage, TRAP‐cage, the disassembly of which can be induced by the addition of reducing agents, is established. In this system, TRAP‐cage with SpyCatcher moieties in the lumen is prepared using genetic modification of the protein building block and assembled into a cage structure with either monovalent gold ions or molecular crosslinkers. The resulting protein cage can efficiently capture guest proteins equipped with a SpyTag by simply mixing them in an aqueous solution. This post‐assembly loading system, which circumvents the exposure of guests to thiol‐reactive crosslinkers, enables the packaging of enzymes possessing a catalytic cysteine or a metal cofactor while retaining their catalytic activity.

中文翻译：

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重新设计人工蛋白笼以有效包装活性酶

易于封装感兴趣的活性酶的蛋白质笼为递送和催化提供了有用的纳米工具，其中具有可编程拆卸特性的蛋白质笼作为特别有吸引力的平台。在这里，建立了一种基于人工蛋白笼（TRAP-cage）的通用客体包装系统，可以通过添加还原剂来诱导其分解。在该系统中，通过对蛋白质构件进行基因修饰来制备内腔中具有 SpyCatcher 部分的 TRAP 笼，并用单价金离子或分子交联剂组装成笼结构。只需将带有 SpyTag 的客体蛋白简单地混合在水溶液中，所得的蛋白笼就可以有效地捕获它们。这种组装后加载系统避免了客体暴露于硫醇反应性交联剂，使得能够包装具有催化半胱氨酸或金属辅因子的酶，同时保留其催化活性。