Analysis of Cytoplasmic and Membrane Molecular Crowding in Genetically Programmed Synthetic Cells.,Biomacromolecules

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Analysis of Cytoplasmic and Membrane Molecular Crowding in Genetically Programmed Synthetic Cells.
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-05-22 , DOI: 10.1021/acs.biomac.0c00513
David Garenne ₁ , Vincent Noireaux ₁

Affiliation

Building genetically programmed synthetic cell systems by molecular integration is a powerful and effective approach to capture the synergies between biomolecules when they are put together. In this work, we characterized quantitatively the effects of molecular crowding on gene expression in the cytoplasm of minimal cells, when a crowding agent is added to the reaction, and on protein self-assembly at the membrane, when a crowding agent is attached to the lipid bilayer. We demonstrate that achieving membrane crowding only is sufficient to keep cytoplasmic expression at its highest and to promote the polymerization of the MreB cytoskeletal protein at the lipid bilayer into a network that is mechanically sturdy. Furthermore, we show that membrane crowding can be emulated by different types of macromolecules, supporting a purely entropic mode of action for supramolecular assembly of cytoskeletal proteins at the bilayer. These unanticipated results provide quantitative and general insights relevant to synthetic cell builders.

中文翻译：

转基因合成细胞中细胞质和膜分子的拥挤分析。

通过分子整合构建基因编程的合成细胞系统是一种强大而有效的方法，可以将生物分子放在一起时捕获它们之间的协同作用。在这项工作中，我们定量表征了分子拥挤对最小细胞细胞质中基因表达的影响（当将拥挤剂添加到反应中时）以及对膜上蛋白质自组装的影响（当拥挤剂附着到膜上时）。脂质双分子层。我们证明，仅实现膜拥挤就足以保持最高的细胞质表达并促进脂质双层的MreB细胞骨架蛋白聚合成机械坚固的网络。此外，我们表明膜拥挤可以通过不同类型的大分子来模拟，支持在双层中细胞骨架蛋白超分子组装的纯熵作用模式。这些出乎意料的结果提供了与合成细胞生成器有关的定量和一般性见解。

更新日期：2020-07-13

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