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Pulsed electric fields induce modulation of protein liquid-liquid phase separation.
Soft Matter ( IF 2.9 ) Pub Date : 2020-09-03 , DOI: 10.1039/d0sm01478h
Mengying Wang 1 , Sven Falke 1 , Robin Schubert 2 , Kristina Lorenzen 2 , Qing-di Cheng 1 , Christian Exner 1 , Hévila Brognaro 1 , Célestin Nzanzu Mudogo 3 , Christian Betzel 1
Affiliation  

The time-resolved dynamic assembly and the structures of protein liquid dense clusters (LDCs) were analyzed under pulsed electric fields (EFs) applying complementary polarized and depolarized dynamic light scattering (DLS/DDLS), optical microscopy, and transmission electron microscopy (TEM). We discovered that pulsed EFs substantially affected overall morphologies and spatial distributions of protein LDCs and microcrystals, and affected the phase diagrams of LDC formation, including enabling protein solutions to overcome the diffusive flux energy barrier to phase separate. Data obtained from DLS/DDLS and TEM showed that LDCs appeared as precursors of protein crystal nuclei, followed by the formation of ordered structures within LDCs applying a pulsed EF. Experimental results of circular dichroism spectroscopy provided evidence that the protein secondary structure content is changing under EFs, which may consequently modulate protein–protein interactions, and the morphology, dimensions, and internal structure of LDCs. Data and results obtained unveil options to modulate the phase diagram of crystallization, and physical morphologies of protein LDCs and microcrystals by irradiating sample suspensions with pulsed EFs.

中文翻译:

脉冲电场引起蛋白质液-液相分离的调节。

利用互补极化和去极化动态光散射(DLS / DDLS),光学显微镜和透射电子显微镜(TEM)在脉冲电场(EFs)下分析了时间分辨的动态组装和蛋白液体密集簇(LDC)的结构。我们发现脉冲EF实质上影响了蛋白质LDC和微晶的整体形态和空间分布,并影响了LDC形成的相图,包括使蛋白质溶液能够克服扩散通量能垒以实现相分离。从DLS / DDLS和TEM获得的数据表明,LDC出现为蛋白质晶体核的前体,随后在LDC内应用脉冲EF形成有序结构。圆二色光谱的实验结果提供了证据,证明在EFs下蛋白质二级结构的含量正在变化,这可能因此调节了蛋白质与蛋白质的相互作用,以及最不发达国家的形态,尺寸和内部结构。获得的数据和结果揭示了通过用脉冲EF辐射样品悬浮液来调节蛋白质LDC和微晶的结晶相图以及物理形态的选项。
更新日期:2020-09-30
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