Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm
Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm
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DOI：
10.1038/nchembio.2535
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发表时间：
2018-02-01
影响因子：
14.8
通讯作者：
Warren, Martin J.
中科院分区：
文献类型：
--
作者：
Lee, Matthew J.;Mantell, Judith;Warren, Martin J.
我们已经开发了一种系统，用于生产渗透到整个大肠杆菌细胞质的超分子支架。这种细胞支架是由一个细菌微室壳蛋白和两个互补的从头开始卷曲的多肽组成的三组分系统构建的。我们表明，其他蛋白质可以靶向这种细胞内的丝状排列。具体地说，丙酮酸脱羧酶和酒精脱氢酶直接作用于细丝，导致这些工程细菌细胞比那些不产生支架的细胞产生更多的乙醇。这与通过酶协同定位提高代谢效率是一致的。最后，壳蛋白支架可以定向到细胞的内膜，展示了如何通过细胞内蛋白设计将合成的细胞组织与空间优化结合在一起。这种细胞支架具有开发下一代细胞工厂的潜力，可以用来组织酶途径和代谢物转运体，以提高代谢流量。
We have developed a system for producing a supramolecular scaffold that permeates the entire Escherichia coli cytoplasm. This cytoscaffold is constructed from a three-component system comprising a bacterial microcompartment shell protein and two complementary de novo coiled-coil peptides. We show that other proteins can be targeted to this intracellular filamentous arrangement. Specifically, the enzymes pyruvate decarboxylase and alcohol dehydrogenase have been directed to the filaments, leading to enhanced ethanol production in these engineered bacterial cells compared to those that do not produce the scaffold. This is consistent with improved metabolic efficiency through enzyme colocation. Finally, the shell-protein scaffold can be directed to the inner membrane of the cell, demonstrating how synthetic cellular organization can be coupled with spatial optimization through in-cell protein design. The cytoscaffold has potential in the development of next-generation cell factories, wherein it could be used to organize enzyme pathways and metabolite transporters to enhance metabolic flux.