DMSO对DNA构象和力学的影响。

IF 3.2 3区 生物学 Q2 BIOPHYSICS
Koen R Storm, Caroline Körösy, Enrico Skoruppa, Stefanie D Pritzl, Pauline J Kolbeck, Willem Vanderlinden, Helmut Schiessel, Jan Lipfert
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引用次数: 0

摘要

二甲基亚砜(DMSO)是一种极性非质子溶剂,用途广泛,包括用作药物和药物输送,作为荧光染料的溶剂,以及在处理DNA的酶促反应中。因此,许多测定含有低浓度(≤10%)的二甲基亚砜。众所周知,DMSO降低了DNA的熔化温度,但其对DNA构象和低于熔化温度的机械性能的影响尚不清楚。在这里,我们使用互补的单分子技术在0-60% DMSO的存在下探测DNA。磁镊子力延伸测量发现,DNA的弯曲持续长度随着DMSO浓度的增加而适度线性减少,达到20 vol%,每%-DMSO减少(0.43±0.02)%。磁镊子扭扭测量表明,DMSO存在时,熔化扭矩减少,发现DNA的螺旋扭扭在20% DMSO浓度下基本保持不变,而更高浓度的DMSO会略微解开螺旋。使用AFM成像,我们发现DMSO对DNA构象的适度压实,并观察到端到端距离的平均平方系统减少1.2% -DMSO。我们使用粗粒度蒙特卡罗模拟DNA作为具有可变密度的柔性段的半柔性聚合物,代表dmso诱导的局部缺陷或熔化,以使观察到的行为合理化。该模型量化了在DNA中引入局部柔性区域的影响,并给出了与磁镊子和原子力显微镜成像实验一致的趋势。我们的研究结果表明,添加高达50%的DMSO对DNA结构和力学的影响是渐进的,而低浓度(≤20%)的诱导变化相对较小。我们的工作为理解和模拟DMSO在一系列生物物理和生化分析中对DNA的影响提供了一个基线。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The effects of DMSO on DNA conformations and mechanics.

Dimethyl sulfoxide (DMSO) is a polar aprotic solvent used in a wide range of applications, including uses as a drug and in drug delivery, as a solvent for fluorescent dyes, and in enzymatic reactions that process DNA. Consequently, many assays contain low concentrations (≤10%) of DMSO. While it is well known that DMSO lowers the melting temperature of DNA, its effects on DNA conformations and mechanical properties below the melting temperature are unclear. Here, we use complementary single-molecule techniques to probe DNA in the presence of 0-60% DMSO. Magnetic tweezers force-extension measurements find that the bending persistence length of DNA decreases moderately and linearly with DMSO concentrations up to 20 vol %, by (0.43 ± 0.02%) per %-DMSO. Magnetic tweezers twist measurements demonstrate a reduction in melting torque in the presence of DMSO and find that the helical twist of DNA remains largely unchanged up to 20% DMSO, while even higher concentrations slightly unwind the helix. Using AFM imaging, we find a moderate compaction of DNA conformations by DMSO and observe a systematic decrease of the mean-squared end-to-end distance by 1.2% per %-DMSO. We use coarse-grained Monte Carlo simulations of DNA as a semiflexible polymer with a variable density of flexible segments, representing DMSO-induced local defects or melting, to rationalize the observed behavior. The model quantitates the effects of introducing locally flexible regions into DNA and gives trends in line with the magnetic tweezers and AFM imaging experiments. Our results show that addition of up to 50% DMSO has a gradual effect on DNA structure and mechanics and that, for low concentrations (≤20%), the induced changes are relatively minor. Our work provides a baseline to understand and model the effects of DMSO on DNA in a range of biophysical and biochemical assays.

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来源期刊
Biophysical journal
Biophysical journal 生物-生物物理
CiteScore
6.10
自引率
5.90%
发文量
3090
审稿时长
2 months
期刊介绍: BJ publishes original articles, letters, and perspectives on important problems in modern biophysics. The papers should be written so as to be of interest to a broad community of biophysicists. BJ welcomes experimental studies that employ quantitative physical approaches for the study of biological systems, including or spanning scales from molecule to whole organism. Experimental studies of a purely descriptive or phenomenological nature, with no theoretical or mechanistic underpinning, are not appropriate for publication in BJ. Theoretical studies should offer new insights into the understanding ofexperimental results or suggest new experimentally testable hypotheses. Articles reporting significant methodological or technological advances, which have potential to open new areas of biophysical investigation, are also suitable for publication in BJ. Papers describing improvements in accuracy or speed of existing methods or extra detail within methods described previously are not suitable for BJ.
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