Programmed spatial organization of biomacromolecules into discrete, coacervate-based protocells.
Programmed spatial organization of biomacromolecules into discrete, coacervate-based protocells.
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DOI：
10.1038/s41467-020-20124-0
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发表时间：
2020-12-08
影响因子：
16.6
通讯作者：
van Hest JCM
中科院分区：
文献类型：
--
作者：
Altenburg WJ;Yewdall NA;Vervoort DFM;van Stevendaal MHME;Mason AF;van Hest JCM
细胞胞质中挤满了高浓度的许多不同的生物大分子，这在自下而上的合成细胞研究中很难模拟，并且限制了现有原细胞平台的功能。因此，显然需要一种通用的、生物相容的且易于使用的工具来更准确地模拟这种环境。在此，我们描述了一种离散的、基于膜结合凝聚层的原细胞平台的开发，该平台利用 Ni2+-次氮基三乙酸和 His 标记蛋白之间众所周知的结合基序，对生物相关大分子的负载进行高水平的控制。该平台可以以受控、高效和良性的方式积累蛋白质，最终增强封装的双酶级联和蛋白酶介导的货物分泌，凸显了该方法的效力。这种对生物相关蛋白质进行程序化空间组织的多功能方法扩展了原细胞工具箱，并为开发下一代复杂但调控良好的合成细胞铺平了道路。模仿合成细胞中拥挤的细胞质一直是其功能的主要限制。在这里，作者利用镍、次氮基三乙酸和组氨酸标记蛋白之间的相互作用来控制大分子加载到空间编程的基于凝聚层的原始细胞中。
The cell cytosol is crowded with high concentrations of many different biomacromolecules, which is difficult to mimic in bottom-up synthetic cell research and limits the functionality of existing protocellular platforms. There is thus a clear need for a general, biocompatible, and accessible tool to more accurately emulate this environment. Herein, we describe the development of a discrete, membrane-bound coacervate-based protocellular platform that utilizes the well-known binding motif between Ni2+-nitrilotriacetic acid and His-tagged proteins to exercise a high level of control over the loading of biologically relevant macromolecules. This platform can accrete proteins in a controlled, efficient, and benign manner, culminating in the enhancement of an encapsulated two-enzyme cascade and protease-mediated cargo secretion, highlighting the potency of this methodology. This versatile approach for programmed spatial organization of biologically relevant proteins expands the protocellular toolbox, and paves the way for the development of the next generation of complex yet well-regulated synthetic cells. Mimicking the crowded cytosol of cells in synthetic cells has been a major limitation to the functionality. Here, the authors used the interaction between nickel, nitrilotriacetic acid and histidine tagged proteins to control loading of macromolecules into spatially programmed coacervate-based protocells.
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影响因子：
21.8
作者：
Adamala KP;Martin-Alarcon DA;Guthrie-Honea KR;Boyden ES
通讯作者：
Boyden ES
影响因子：
21.8
作者：
Kurihara, Kensuke;Tamura, Mieko;Sugawara, Tadashi
通讯作者：
Sugawara, Tadashi
影响因子：
16.6
作者：
Deshpande, Siddharth;Brandenburg, Frank;Dekker, Cees
通讯作者：
Dekker, Cees
影响因子：
46.9
作者：
Castellana M;Wilson MZ;Xu Y;Joshi P;Cristea IM;Rabinowitz JD;Gitai Z;Wingreen NS
通讯作者：
Wingreen NS
DOI：
10.1126/science.1198701
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Good MC;Zalatan JG;Lim WA
通讯作者：
Lim WA