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电场作用于金字塔结构对酶生物分子影响的原子力显微镜研究

Atomic Force Microscopy Study of the Effect of an Electric Field, Applied to a Pyramidal Structure, on Enzyme Biomolecules.

作者信息

Ivanov Yuri D, Tatur Vadim Y, Shumov Ivan D, Kozlov Andrey F, Valueva Anastasia A, Ivanova Irina A, Ershova Maria O, Ivanova Nina D, Stepanov Igor N, Lukyanitsa Andrei A, Ziborov Vadim S

机构信息

Institute of Biomedical Chemistry, 119121 Moscow, Russia.

Joint Institute for High Temperatures of the Russian Academy of Sciences, 125412 Moscow, Russia.

出版信息

J Funct Biomater. 2022 Nov 10;13(4):234. doi: 10.3390/jfb13040234.

Abstract

The influence of an external constant strong electric field, formed using a pyramidal structure under a high electric potential, on an enzyme located near its apex, is studied. Horseradish peroxidase (HRP) is used as a model. In our experiments, a 27 kV direct current (DC) voltage was applied to two electrodes with a conducting pyramidal structure attached to one of them. The enzyme particles were visualized by atomic force microscopy (AFM) after the adsorption of the enzyme from its 0.1 µM solution onto mica AFM substrates. It is demonstrated that after the 40 min exposure to the electric field, the enzyme forms extended structures on mica, while in control experiments compact HRP particles are observed. After the exposure to the electric field, the majority of mica-adsorbed HRP particles had a height of 1.2 nm (as opposed to 1.0 nm in the case of control experiments), and the contribution of higher (>2.0 nm) particles was also considerable. This indicates the formation of high-order HRP aggregates under the influence of an applied electric field. At that, the enzymatic activity of HRP against its substrate 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) remains unaffected. These results are important for studying macroscopic effects of strong electromagnetic fields on enzymes, as well as for the development of cellular structure models.

摘要

研究了利用处于高电势下的金字塔结构形成的外部恒定强电场对位于其顶点附近的酶的影响。以辣根过氧化物酶(HRP)作为模型。在我们的实验中,将27 kV直流(DC)电压施加到两个电极上,其中一个电极附着有导电金字塔结构。在将酶从其0.1 µM溶液吸附到云母原子力显微镜(AFM)基底上之后,通过原子力显微镜观察酶颗粒。结果表明,在电场中暴露40分钟后,酶在云母上形成了伸展结构,而在对照实验中观察到的是紧密的HRP颗粒。在电场作用后,大多数云母吸附的HRP颗粒高度为1.2 nm(与对照实验中的1.0 nm相反),并且更高(>2.0 nm)颗粒的贡献也相当可观。这表明在施加电场的影响下形成了高阶HRP聚集体。此时,HRP对其底物2,2′-叠氮基双(3-乙基苯并噻唑啉-6-磺酸盐)(ABTS)的酶活性保持不受影响。这些结果对于研究强电磁场对酶的宏观效应以及细胞结构模型的开发具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/958b/9680214/c39f4efe4512/jfb-13-00234-g001.jpg

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