Intrinsically disordered protein biosensor tracks the physical-chemical effects of osmotic stress on cells.
Intrinsically disordered protein biosensor tracks the physical-chemical effects of osmotic stress on cells.
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DOI：
10.1038/s41467-021-25736-8
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发表时间：
2021-09-14
影响因子：
16.6
通讯作者：
Dinneny JR
中科院分区：
文献类型：
--
作者：
Cuevas-Velazquez CL;Vellosillo T;Guadalupe K;Schmidt HB;Yu F;Moses D;Brophy JAN;Cosio-Acosta D;Das A;Wang L;Jones AM;Covarrubias AA;Sukenik S;Dinneny JR
当外部环境的剧烈变化导致细胞内部的物理化学性质发生变化时，细胞的动态平衡就会受到干扰。目前尚缺乏动态监测这些细胞内效应的实验方法。在这里，我们利用固有无序蛋白区域(IDR)的环境敏感性和结构可塑性来开发一种FRET生物传感器，能够监测渗透胁迫引起的细胞内快速变化。这种名为SED1的生物传感器利用了拟南芥在缺水条件下表达的内在无序的AtLEA4-5蛋白。计算模型和体外研究表明，SED1对大分子拥挤高度敏感。SED1在活的细菌、酵母、植物和人类细胞中显示出大的和近线性的渗透压依赖的FRET变化，证明了这种工具在研究与水相关的压力方面的广泛用途。这项研究展示了IDR感知生命之树上细胞环境的非凡能力，并为将其用作环境响应型分子工具提供了蓝图。目前缺乏监测细胞渗透压依赖性变化的方法。在这里，作者利用在植物缺水条件下表达的拟南芥内在无序的AtLEA4-5蛋白来开发一种FRET生物传感器(SED1)来监测渗透胁迫。
Cell homeostasis is perturbed when dramatic shifts in the external environment cause the physical-chemical properties inside the cell to change. Experimental approaches for dynamically monitoring these intracellular effects are currently lacking. Here, we leverage the environmental sensitivity and structural plasticity of intrinsically disordered protein regions (IDRs) to develop a FRET biosensor capable of monitoring rapid intracellular changes caused by osmotic stress. The biosensor, named SED1, utilizes the Arabidopsis intrinsically disordered AtLEA4-5 protein expressed in plants under water deficit. Computational modeling and in vitro studies reveal that SED1 is highly sensitive to macromolecular crowding. SED1 exhibits large and near-linear osmolarity-dependent changes in FRET inside living bacteria, yeast, plant, and human cells, demonstrating the broad utility of this tool for studying water-associated stress. This study demonstrates the remarkable ability of IDRs to sense the cellular environment across the tree of life and provides a blueprint for their use as environmentally-responsive molecular tools. Methods to monitor osmolarity-dependent changes in cell are currently lacking. Here the authors use the Arabidopsis intrinsically disordered AtLEA4-5 protein, which is expressed in plants under water deficit, to develop a FRET biosensor (SED1) to monitor osmotic stress.
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影响因子：
--
作者：
Gnutt, David;Brylski, Oliver;Ebbinghaus, Simon
通讯作者：
Ebbinghaus, Simon
影响因子：
3.4
作者：
McGibbon, Robert T.;Beauchamp, Kyle A.;Pande, Vijay S.
通讯作者：
Pande, Vijay S.
影响因子：
7.7
作者：
Jones AM;Danielson JA;Manojkumar SN;Lanquar V;Grossmann G;Frommer WB
通讯作者：
Frommer WB
DOI：
10.1085/jgp.201411295
发表时间：
2015-05
期刊：
The Journal of general physiology
影响因子：
--
作者：
Haswell ES;Verslues PE
通讯作者：
Verslues PE
影响因子：
64.8
作者：
Keul, Nicholas D.;Oruganty, Krishnadev;Wood, Zachary A.
通讯作者：
Wood, Zachary A.