Exploring the role of preferential solvation in the stability of globular proteins through the study of ovalbumin interaction with organic additives

Tatyana Tretyakova; Maya Makharadze; Sopio Uchaneishvili; Mikhael Shushanyan; Dimitri Khoshtariya; Tatyana Tretyakova; Maya Makharadze; Sopio Uchaneishvili; Mikhael Shushanyan; Dimitri Khoshtariya

doi:10.3934/biophy.2023025

AIMS Biophysics

2023, Volume 10, Issue 4: 440-452. doi: 10.3934/biophy.2023025

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Research article

Exploring the role of preferential solvation in the stability of globular proteins through the study of ovalbumin interaction with organic additives

1.
Laboratory of Biophysics, LEPL I. Beritashvili Center of Experimental Biomedicine, 0160 Tbilisi, Georgia
2.
Institute for Biophysics and Bionanosciences, Faculty of Exact and Natural Sciences, Ivane Javakhishvili Tbilisi State University, 0128 Tbilisi, Georgia

Received: 30 May 2023 Revised: 24 September 2023 Accepted: 10 October 2023 Published: 25 October 2023

The impact of denaturing and stabilizing osmolytes on protein conformational dynamics has been extensively explored due to the significant contribution of protein solvation to the stability, function, malfunction and regulation of globular proteins. We studied the effect of two nonspecific organic molecules, urea, which is a conventional denaturant, and dimethyl sulfoxide (DMSO), which is a multilateral organic solvent, on the stability and conformational dynamics of a non-inhibitory serpin, ovalbumin (OVA). A differential scanning microcalorimetry (DSC) experimental series conducted in the phosphate buffer solutions containing 0–30% of additives revealed the destabilizing impact of both urea and DMSO in a mild acidic media, manifested in the gradual decrease of thermal unfolding enthalpy and transition temperature. These findings differ from the results observed in our study of the mild alkaline DMSO buffered solutions of OVA, where the moderate stabilization of OVA was observed in presence of 5–10% of DMSO. However, the overall OVA interaction patterns with urea and DMSO are consistent with our previous findings on the stability and conformational flexibility of another model globular protein, α-chymotrypsin, in similar medium conditions. The obtained results could be explained by preferential solvation patterns. Positive preferential solvation of protein by urea in urea/water mixtures mainly weakens the hydrophobic interactions of the protein globule and eventually leads to the disruption of the tertiary structure within the whole range of urea concentrations. Alternatively, under certain experimental conditions in DMSO/water mixtures, positive preferential solvation by water molecules can be observed. We assume that the switch to the positive preferential solvation by DMSO, which is shown to have a soft maximum around 20–30% DMSO, could be shifted towards lower additive concentrations due to the intrinsic capability of ovalbumin OVA to convert into a heat-stable, yet flexible set of conformations that have increased the surface hydrophobicity, characteristic to molten-globule-like states.
- globular protein,
- protein stability,
- thermal unfolding,
- differential scanning calorimetry,
- ovalbumin,
- urea,
- dimethyl sulfoxide
Citation: Tatyana Tretyakova, Maya Makharadze, Sopio Uchaneishvili, Mikhael Shushanyan, Dimitri Khoshtariya. Exploring the role of preferential solvation in the stability of globular proteins through the study of ovalbumin interaction with organic additives[J]. AIMS Biophysics, 2023, 10(4): 440-452. doi: 10.3934/biophy.2023025

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Abstract

The impact of denaturing and stabilizing osmolytes on protein conformational dynamics has been extensively explored due to the significant contribution of protein solvation to the stability, function, malfunction and regulation of globular proteins. We studied the effect of two nonspecific organic molecules, urea, which is a conventional denaturant, and dimethyl sulfoxide (DMSO), which is a multilateral organic solvent, on the stability and conformational dynamics of a non-inhibitory serpin, ovalbumin (OVA). A differential scanning microcalorimetry (DSC) experimental series conducted in the phosphate buffer solutions containing 0–30% of additives revealed the destabilizing impact of both urea and DMSO in a mild acidic media, manifested in the gradual decrease of thermal unfolding enthalpy and transition temperature. These findings differ from the results observed in our study of the mild alkaline DMSO buffered solutions of OVA, where the moderate stabilization of OVA was observed in presence of 5–10% of DMSO. However, the overall OVA interaction patterns with urea and DMSO are consistent with our previous findings on the stability and conformational flexibility of another model globular protein, α-chymotrypsin, in similar medium conditions. The obtained results could be explained by preferential solvation patterns. Positive preferential solvation of protein by urea in urea/water mixtures mainly weakens the hydrophobic interactions of the protein globule and eventually leads to the disruption of the tertiary structure within the whole range of urea concentrations. Alternatively, under certain experimental conditions in DMSO/water mixtures, positive preferential solvation by water molecules can be observed. We assume that the switch to the positive preferential solvation by DMSO, which is shown to have a soft maximum around 20–30% DMSO, could be shifted towards lower additive concentrations due to the intrinsic capability of ovalbumin OVA to convert into a heat-stable, yet flexible set of conformations that have increased the surface hydrophobicity, characteristic to molten-globule-like states.

Abbreviations

DSC

differential scanning calorimetry

OVA

ovalbumin

S-OVA

S-ovalbumin

DMSO

dimethyl sulfoxide

α-CT

α-chymotrypsin

Acknowledgments

This work was supported by Shota Rustaveli National Science Foundation of Georgia (SRNSFG) research grant for young scientists YS-18-2034.

Conflict of interest

The authors declare no conflict of interest.

References

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