Programmed loading and rapid purification of engineered bacterial microcompartment shells
Programmed loading and rapid purification of engineered bacterial microcompartment shells
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DOI：
10.1038/s41467-018-05162-z
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发表时间：
2018-07-23
影响因子：
16.6
通讯作者：
Kerfeld, Cheryl A.
中科院分区：
文献类型：
--
作者：
Hagen, Andrew;Sutter, Markus;Kerfeld, Cheryl A.
细菌微室(BMC)是一种选择性可渗透的蛋白质细胞器，它包裹着跨越细菌门的代谢途径的片段。它们由一个酶核心和一个由多个不同蛋白质组成的蛋白质壳组成。尽管在各种生物技术应用中具有巨大的潜力，但由于分离和表征它们的困难，以及缺乏编程岩芯和壳层渗透率的可靠方法，工程努力一直受到阻碍。我们通过使用亲和手柄使壳蛋白功能化来解决这些挑战，使基于互补的亲和纯化(CAP)和特定的货物对接位置能够通过共价键(Enco)进行有效的封装。这些外壳功能化扩展了我们对BMC架构原则的了解，并使开发精确定义结构和组成的最小外壳系统成为可能。CAP和Enco的通用性将使它们能够应用于不同功能的微室系统，以便于表征自然功能和开发用于选择性划分蛋白质的定制壳。
Bacterial microcompartments (BMCs) are selectively permeable proteinaceous organelles which encapsulate segments of metabolic pathways across bacterial phyla. They consist of an enzymatic core surrounded by a protein shell composed of multiple distinct proteins. Despite great potential in varied biotechnological applications, engineering efforts have been stymied by difficulties in their isolation and characterization and a dearth of robust methods for programming cores and shell permeability. We address these challenges by functionalizing shell proteins with affinity handles, enabling facile complementation-based affinity purification (CAP) and specific cargo docking sites for efficient encapsulation via covalent-linkage (EnCo). These shell functionalizations extend our knowledge of BMC architectural principles and enable the development of minimal shell systems of precisely defined structure and composition. The generalizability of CAP and EnCo will enable their application to functionally diverse microcompartment systems to facilitate both characterization of natural functions and the development of bespoke shells for selectively compartmentalizing proteins.