Investigation of the Influence of Liquid Motion in a Flow-based System on an Enzyme Aggregation State with an Atomic Force Microscopy Sensor: The Effect of Water Flow

Ziborov, Vadim S.; Pleshakova, Tatyana O.; Shumov, Ivan D.; Kozlov, Andrey F.; Ivanova, Irina A.; Valueva, Anastasia A.; Tatur, Vadim Yu.; Negodailov, Andrey N.; Lukyanitsa, Andrei A.; Ivanov, Yuri D.

doi:10.3390/app10134560

Cited by 17 publications

(68 citation statements)

References 32 publications

(60 reference statements)

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“…This fact indicates that the flow-induced field only impacts the spatial structures of the protein globule near its surface, while not affecting its active site. The effects of HRP aggregation in the presence of electromagnetic fields were previously reported in our recent studies [ 8 , 35 , 36 ] and by Sun et al [ 37 , 38 ].…”

Section: Discussionsupporting

confidence: 69%

“…The observed effect of the water, flowing through the coiled silicone pipe, on the adsorbability of HRP is apparently explained by the electromagnetic nature of the flow-induced field. The induction of such an electromagnetic field obviously occurs owing to a triboelectric effect consisting of a generation of charge in the liquid, flowing along a polymeric surface [ 8 , 29 , 30 , 31 , 32 , 33 , 34 ]. The triboelectrically induced charges induce an electromagnetic field, which, in turn, affects the protein during the incubation of its solution near the coil.…”

Section: Discussionmentioning

confidence: 99%

“…Experimental setup, which is analogous to the one used in our previous study [ 8 ], is schematically shown in Figure 1 (see the Introduction). The coil, through which water was pumped at a speed of 4 m/s, was represented by a silicone pipe (outer diameter 10 mm), spiral-wound (13 turns) onto a 170 mm diameter glass cylinder, so that the coil length was 130 mm.…”

Section: Methodsmentioning

confidence: 99%

“…The AFM experiments were carried out using the direct surface adsorption method [ 8 , 15 , 35 , 36 ], similar to [ 8 ]. Briefly, muscovite mica sheets (SPI, West Chester, PA, USA) were used as AFM substrates.…”

Section: Methodsmentioning

confidence: 99%

“…This is often realized by employing flow-based heat exchangers [ 7 ], such as coils, where water is used as a heat-transfer agent. The flow of water through a coiled polymeric pipe was shown to induce an electromagnetic field, which can influence protein molecules, causing changes in their physicochemical properties [ 8 ]. Such a field, induced by liquid flow owing to a triboelectric effect, can extend to outside the coil.…”

Section: Introductionmentioning

confidence: 99%

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AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase

et al. 2021

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Atomic force microscopy (AFM)-based fishing is a promising method for the detection of low-abundant proteins. This method is based on the capturing of the target proteins from the analyzed solution onto a solid substrate, with subsequent counting of the captured protein molecules on the substrate surface by AFM. Protein adsorption onto the substrate surface represents one of the key factors determining the capturing efficiency. Accordingly, studying the factors influencing the protein adsorbability onto the substrate surface represents an actual direction in biomedical research. Herein, the influence of water motion in a flow-based system on the protein adsorbability and on its enzymatic activity has been studied with an example of horseradish peroxidase (HRP) enzyme by AFM, attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) and conventional spectrophotometry. In the experiments, HRP solution was incubated in a setup modeling the flow section of a biosensor communication. The measuring cell with the protein solution was placed near a coiled silicone pipe, through which water was pumped. The adsorbability of the protein onto the surface of the mica substrate has been studied by AFM. It has been demonstrated that incubation of the HRP solution near the coiled silicone pipe with flowing water leads to an increase in its adsorbability onto mica. This is accompanied by a change in the enzyme’s secondary structure, as has been revealed by ATR-FTIR. At the same time, its enzymatic activity remains unchanged. The results reported herein can be useful in the development of models describing the influence of liquid flow on the properties of enzymes and other proteins. The latter is particularly important for the development of biosensors for biomedical applications—particularly for serological analysis, which is intended for the early diagnosis of various types of cancer and infectious diseases. Our results should also be taken into account in studies of the effects of protein aggregation on hemodynamics, which plays a key role in human body functioning.

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Section: Discussionsupporting

confidence: 69%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase

et al. 2021

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AFM study of changes in properties of horseradish peroxidase after incubation of its solution near a pyramidal structure

Ivanov

Pleshakova

Shumov

et al. 2021

Sci Rep

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In our present paper, the influence of a pyramidal structure on physicochemical properties of a protein in buffer solution has been studied. The pyramidal structure employed herein was similar to those produced industrially for anechoic chambers. Pyramidal structures are also used as elements of biosensors. Herein, horseradish peroxidase (HRP) enzyme was used as a model protein. HRP macromolecules were adsorbed from their solution onto an atomically smooth mica substrate, and then visualized by atomic force microscopy (AFM). In parallel, the enzymatic activity of HRP was estimated by conventional spectrophotometry. Additionally, attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) has been employed in order to find out whether or not the protein secondary structure changes after the incubation of its solution either near the apex of a pyramid or in the center of its base. Using AFM, we have demonstrated that the incubation of the protein solution either in the vicinity of the pyramid’s apex or in the center of its base influences the physicochemical properties of the protein macromolecules. Namely, the incubation of the HRP solution in the vicinity of the top of the pyramidal structure has been shown to lead to an increase in the efficiency of the HRP adsorption onto mica. Moreover, after the incubation of the HRP solution either near the top of the pyramid or in the center of its base, the HRP macromolecules adsorb onto the mica surface predominantly in monomeric form. At that, the enzymatic activity of HRP does not change. The results of our present study are useful to be taken into account in the development of novel biosensor devices (including those for the diagnosis of cancer in humans), in which pyramidal structures are employed as sensor, noise suppression or construction elements.

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The Influence of Glycerol Flow through a Shielded Coil on an Enzyme Placed Near Its Side

Ivanov¹,

Shumov²,

Kozlov³

et al. 2023

Preprint

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Glycerol has found its applications as a heat-transfer fluid in heat exchangers, and as a compo-nent of functional solutions in biosensor analysis. Flowing non-aqueous fluids are known to be able to induce electromagnetic fields due to triboelectric effect. These triboelectrically generated electromagnetic fields can affect biological macromolecules. Horseradish peroxidase (HRP) is widely employed as a convenient model object for studying how external electric, magnetic, and electromagnetic fields affect enzymes. Herein, we have studied whether the flow of glycerol in a ground-shielded cylindrical coil affects the HRP enzyme incubated at a 2 cm distance near the coil’s side. Atomic force microscopy (AFM) has been employed in order to study the effect of glycerol flow on HRP at the nanoscale. Increased aggregation of HRP on mica has been observed after the incubation of the enzyme near the coil. Moreover, the enzymatic activity of HRP has also been affected. The results reported can find their application in biotechnology, food technology and life sciences application, considering the development of triboelectric generators, en-zyme-based biosensors and bioreactors with surface-immobilized enzymes. Our data can also be of interest for scientists studying triboelectric phenomena.

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Investigation of the Influence of Liquid Motion in a Flow-based System on an Enzyme Aggregation State with an Atomic Force Microscopy Sensor: The Effect of Water Flow

Cited by 17 publications

References 32 publications

AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase

AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase

AFM study of changes in properties of horseradish peroxidase after incubation of its solution near a pyramidal structure

The Influence of Glycerol Flow through a Shielded Coil on an Enzyme Placed Near Its Side

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