Modulation of global stability, ligand binding and catalytic properties of trypsin by anions

Dušeková, Eva; Garajová, Katarína; Yavaşer, Rukiye; Tomková, Mária; Sedláková, Dagmar; Veronika, Dzurillová; Кулик, Н. В.; Fadaei, Fatemeh; Shaposhnikova, Anastasiia; Minofar, Babak; Sedlák, Erik

doi:10.1016/j.bpc.2022.106856

Cited by 3 publications

(3 citation statements)

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“…The observed slopes of thermodynamic parameters obtained for different salts (e.g., ΔΔH/Δ[salt]) are specific for a given chloride salt and may be correlated with some of the physicochemical properties of the cation. Similar correlations were found in different studies of proteins in the presence of salts of the Hofmeister series [ 42 , 43 , 44 , 45 ]. In our case, we could not find any evidence for a robust correlation between the salt-specific values of the slopes and ion properties such as hydration entropy, hydration number, charge density, polarizability, and viscosity B coefficient (see Figures S6–S17 , Supplementary Materials , which includes also pairwise correlations between thermodynamic parameters of domains).…”

Section: Discussionsupporting

confidence: 88%

Salt-Specific Suppression of the Cold Denaturation of Thermophilic Multidomain Initiation Factor 2

Džupponová

Tomášková

Antošová

et al. 2023

IJMS

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Thermophilic proteins and enzymes are attractive for use in industrial applications due to their resistance against heat and denaturants. Here, we report on a thermophilic protein that is stable at high temperatures (Ttrs, hot 67 °C) but undergoes significant unfolding at room temperature due to cold denaturation. Little is known about the cold denaturation of thermophilic proteins, although it can significantly limit their applications. We investigated the cold denaturation of thermophilic multidomain protein translation initiation factor 2 (IF2) from Thermus thermophilus. IF2 is a GTPase that binds to ribosomal subunits and initiator fMet-tRNAfMet during the initiation of protein biosynthesis. In the presence of 9 M urea, measurements in the far-UV region by circular dichroism were used to capture details about the secondary structure of full-length IF2 protein and its domains during cold and hot denaturation. Cold denaturation can be suppressed by salt, depending on the type, due to the decreased heat capacity. Thermodynamic analysis and mathematical modeling of the denaturation process showed that salts reduce the cooperativity of denaturation of the IF2 domains, which might be associated with the high frustration between domains. This characteristic of high interdomain frustration may be the key to satisfying numerous diverse contacts with ribosomal subunits, translation factors, and tRNA.

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Section: Discussionsupporting

confidence: 88%

Salt-Specific Suppression of the Cold Denaturation of Thermophilic Multidomain Initiation Factor 2

Džupponová

Tomášková

Antošová

et al. 2023

IJMS

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“…To determine whether there is a correlation between ion properties and their efficiency in affecting the rate of the eADF4(C16) fibril elongation, we plotted the slope of dk + /d[salt] from Figure 5a, within the concertation range of 0-150 mM salt, against selected intrinsic properties of the anions. Many parameters reflect different anion properties, but as we pointed out in our previous work, some of these parameters correlate with each other, significantly reducing the number of meaningful dependencies (Dušekov a et al, 2022). As a result, we plotted the parameter dk + /d[salt] versus charge density (Figure 5b) (which also correlates with surface tension and air/water partition coefficient of anions), polarizability (Figure 5d), and viscosity B-coefficient (which also correlates with partition coefficient at hydrocarbon surface) (Figure 5c).…”

Section: Morphology Of Nanofibrilsmentioning

confidence: 90%

“…In 1888, Franz Hofmeister divided ions into kosmotropes and chaotropes based on their capacity to either “fit into” or “break” the ordered structure of water, and their capability to salt out or salt in proteins, respectively (Hofmeister, 1888). By arranging selected anions according to their impact on various protein properties, such as solubility, stability, or activity (Baldwin, 1996; Cacace et al, 1997; Dušeková et al, 2022; Sedlak et al, 2008), the so‐called Hofmeister series of anions can be arranged in the following order:

{normalSO}_{4}^{2 -} > \begin{matrix} lefttrue \\ {HPO}_{4}^{2 -} > {normalF}^{-} > {normalCl}^{-} > {normalBr}^{-} > {normalI}^{-} > {NO}_{3}^{-} \\ > {ClO}_{4}^{-} > {normalSCN}^{-} . \end{matrix}

…”

Section: Introductionmentioning

confidence: 99%

Only kosmotrope anions trigger fibrillization of the recombinant core spidroin eADF4(C16) from Araneus diadematus

Hovanová,

Hovan,

Humenik

et al. 2023

Protein Science

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Recombinant core spidroin eADF4(C16) has received increasing attention due to its ability to form micro‐ and nano‐structured scaffolds, which are based on nanofibrils with great potential for biomedical and biotechnological applications. Phosphate anions have been demonstrated to trigger the eADF4(C16) self‐assembly into cross‐beta fibrils. In the present work, we systematically addressed the effect of nine sodium anions, namely SO42−, HPO42− (Pi), F−, Cl−, Br−, NO3−, I−, SCN−, and ClO4− from the Hofmeister series on the in vitro self‐assembly kinetics of eADF4(C16). We show that besides the phosphate anions, only kosmotropic anions such as sulfate and fluoride can initiate the eADF4(C16) fibril formation. Global analysis of the self‐assembly kinetics, utilizing the platform AmyloFit, showed the nucleation‐based mechanism with a major role of a secondary nucleation, surprisingly independent of the type of the kosmotropic anion. The rate constant of the fibril elongation in mixtures of phosphate anions with other studied anions correlated with their kosmotropic or chaotropic position in the Hofmeister series. Our findings suggest an important role of anion hydration in the eADF4(C16) fibrillization process.This article is protected by copyright. All rights reserved.

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Trypsin-Based Chemoenzymatic Assay for Detection of Pollutants and Safety Assessment of Food Additives

et al. 2023

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Chemoenzymatic assay systems are widely used to detect toxicants in various samples, including food and environment specimens. These methods are based on the ability of various types of toxicant to specifically inhibit/activate the functions of individual enzymes or enzyme systems. The present study examines the possibility of using the proteolytic enzyme trypsin as a specific marker to detect protease inhibitors in different samples. The study shows that trypsin activity is not affected by various heavy metals, pesticides, or quinones at levels considerably greater than their maximum allowable concentrations (MACs) in water bodies. At the same time, the IC50 value for the food preservative potassium sorbate (E202) is 15 mg/L, which is substantially lower than its acceptable daily intake (ADI). The quenching of trypsin fluorescence in the presence of potassium sorbate suggests that inhibition could occur due to the binding of the preservative to the enzyme in the region adjacent to the active center. The trypsin was immobilized in starch gel to ensure its stability in the enzyme inhibition based assay. Single-use reagents were prepared as dry starch disks that could be stored over long periods. Their sensitivity to copper (II) chloride, potassium sorbate, and chromium (III) chloride was similar to the sensitivity of the free trypsin.

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Modulation of global stability, ligand binding and catalytic properties of trypsin by anions

Cited by 3 publications

References 84 publications

Salt-Specific Suppression of the Cold Denaturation of Thermophilic Multidomain Initiation Factor 2

Salt-Specific Suppression of the Cold Denaturation of Thermophilic Multidomain Initiation Factor 2

Only kosmotrope anions trigger fibrillization of the recombinant core spidroin eADF4(C16) from Araneus diadematus

Trypsin-Based Chemoenzymatic Assay for Detection of Pollutants and Safety Assessment of Food Additives

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