De novo design of protein logic gates
De novo design of protein logic gates
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DOI：
10.1126/science.aay2790
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发表时间：
2020-04-03
期刊：
影响因子：
56.9
通讯作者：
Baker, David
中科院分区：
文献类型：
--
作者：
Chen, Zibo;Kibler, Ryan D.;Baker, David
在转录后水平设计用于调节蛋白质功能的模块化蛋白质逻辑是合成生物学面临的一个挑战。在此，我们描述了由从头设计的蛋白质构建的双输入“与”门、“或”门、“与非”门、“或非”门、“同或”门和“非”门的设计。这些门在体外、酵母以及原代人T细胞中调节从分裂酶到转录机制等任意蛋白质单元的结合，它们控制与T细胞耗竭相关的TIM3基因的表达。通过天然质谱法确认的设计结合相互作用协同性使得这些门在很大程度上对输入的化学计量失衡不敏感，并且该方法的模块性能够容易地扩展到三输入“或”门、“与”门和析取范式门。控制元件的模块性和协同性，加上从头设计几乎无限数量蛋白质组分的能力，应该能够实现对广泛生物功能的复杂翻译后控制逻辑的设计。
The design of modular protein logic for regulating protein function at the posttranscriptional level is a challenge for synthetic biology. Here, we describe the design of two-input AND, OR, NAND, NOR, XNOR, and NOT gates built from de novo-designed proteins. These gates regulate the association of arbitrary protein units ranging from split enzymes to transcriptional machinery in vitro, in yeast and in primary human T cells, where they control the expression of the TIM3 gene related to T cell exhaustion. Designed binding interaction cooperativity, confirmed by native mass spectrometry, makes the gates largely insensitive to stoichiometric imbalances in the inputs, and the modularity of the approach enables ready extension to three-input OR, AND, and disjunctive normal form gates. The modularity and cooperativity of the control elements, coupled with the ability to de novo design an essentially unlimited number of protein components, should enable the design of sophisticated posttranslational control logic over a wide range of biological functions.