Diversification of Protein Cage Structure Using Circularly Permuted Subunits
Diversification of Protein Cage Structure Using Circularly Permuted Subunits
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DOI：
10.1021/jacs.7b10513
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发表时间：
2018-01-17
影响因子：
15
通讯作者：
Hilvert, Donald
中科院分区：
文献类型：
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作者：
Azuma, Yusuke;Herger, Michael;Hilvert, Donald
自组装蛋白质笼可用作纳米级分子容器，用于生物技术和医学的各种应用。为了扩大这种系统的效用，人们对定制天然笼形成蛋白的结构和设计新的结构非常感兴趣。在这里，我们报告了Lumazine合酶（一种来自风产液菌（AaLS）的笼形成酶）的循环排列变体，为构建纳米隔室提供了多功能的构建模块，可以轻松生产、定制和多样化。拓扑改变的蛋白质cpAaLS自组装成球形和管状笼状结构，其形态可以通过连接天然末端的接头的长度来控制。此外，cpAaLS蛋白整合到野生型和其他工程AaLS组件：通过在大肠杆菌中共同生产，形成拼凑的笼子。这种共组装策略使得能够将客体蛋白包封在管腔中，通过基因融合修饰外部，以及调整隔室的大小和静电。这除了家庭的AaLS笼拓宽了该系统的范围，为进一步的应用，并突出了实用性的循环置换作为一个潜在的一般策略，用于定制的笼形成蛋白质的性质。
Self-assembling protein cages are useful as nanoscale molecular containers for diverse applications in biotechnology and medicine. To expand the utility of such systems, there is considerable interest in customizing the structures of natural cage-forming proteins and designing new ones. Here we report that a circularly permuted variant of lumazine synthase, a cage-forming enzyme from Aquifex aeolicus (AaLS) affords versatile building blocks for the construction of nano compartments that can be easily produced, tailored, and diversified. The topologically altered protein, cpAaLS, self-assembles into spherical and tubular cage structures with morphologies that can be controlled by the length of the linker connecting the native termini. Moreover, cpAaLS proteins integrate into wild-type and other engineered AaLS assemblies : by coproduction in Escherichia coli to form patchwork cages. This coassembly strategy enables encapsulation of guest proteins in the lumen, modification of the exterior through genetic fusion, and tuning of the size and electrostatics of the compartments. This addition to the family of AaLS cages broadens the scope of this system for further applications and highlights the utility of circular permutation as a potentially general strategy for tailoring the properties of cage-forming proteins.