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共溶质埃克托因对DNA电荷中和的促进与抑制作用

The promotion and suppression of DNA charge neutralization by the cosolute ectoine.

作者信息

Chen Benteng, Wang Yanwei, Yang Guangcan

机构信息

Department of Physics, Wenzhou University Wenzhou 325035 China

出版信息

RSC Adv. 2019 Dec 12;9(70):41050-41057. doi: 10.1039/c9ra09355a. eCollection 2019 Dec 9.

Abstract

Ectoine, a cosolute and osmolyte, is used by extremophilic microorganisms to maintain an osmotic equilibrium of cells with their surrounding medium under conditions of extreme salinity or thermal and pressure stresses. It is also considered a protectant of biomolecules such as protein and DNA in cells. In the present study, we investigate its influence on DNA charge neutralization and compaction through dynamic light scattering (DLS), atomic force microscopy (AFM) and single molecular magnetic tweezers (MT). We found that ectoine can promote DNA charge neutralization induced by multivalent cations at mild cosolute concentration in solution. When the concentration of ectoine is high enough, however, a mixed effect of promotion and suppression can be found under the same ionic conditions. In this case, the electrophoretic mobility (EM) of DNA is promoted in the region of low cation concentration, while suppressed in the region of high counterionic concentration. The charge neutralization of DNA by ectoine is also related to DNA compaction. The promotion and suppression of DNA compaction by ectoine was observed by AFM imaging. The condensed structure of DNA becomes more compact and then loose once more with the increasing concentration of ectoine. Meanwhile, the condensing forces of DNA measured by magnetic tweezers shows the same trend as does the DNA EM. We explained the experimental findings through the combined effect of two intrinsic features of ectoine, preferential exclusion and enhancement of the dielectric constant of the medium.

摘要

四氢嘧啶作为一种共溶质和渗透溶质,被嗜极微生物用于在极端盐度、热应激和压力应激条件下维持细胞与其周围介质的渗透平衡。它也被认为是细胞中蛋白质和DNA等生物分子的保护剂。在本研究中,我们通过动态光散射(DLS)、原子力显微镜(AFM)和单分子磁镊(MT)研究了其对DNA电荷中和及压缩的影响。我们发现,在溶液中适度的共溶质浓度下,四氢嘧啶可以促进多价阳离子诱导的DNA电荷中和。然而,当四氢嘧啶的浓度足够高时,在相同的离子条件下会发现促进和抑制的混合效应。在这种情况下,DNA的电泳迁移率(EM)在低阳离子浓度区域被促进,而在高抗衡离子浓度区域被抑制。四氢嘧啶对DNA的电荷中和也与DNA压缩有关。通过AFM成像观察到了四氢嘧啶对DNA压缩的促进和抑制作用。随着四氢嘧啶浓度的增加,DNA的凝聚结构变得更加紧凑,然后又再次变得松散。同时,用磁镊测量的DNA凝聚力与DNA的EM呈现相同的趋势。我们通过四氢嘧啶的两个固有特性——优先排斥和介质介电常数的增强的综合作用来解释实验结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/308e/9076405/ca37ae627da1/c9ra09355a-f1.jpg

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