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Design and Analysis of a Mutant form of the Ice-Binding Protein from Choristoneura fumiferana

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Abstract

Ice-binding proteins are expressed in the cells of some cold adapted organisms, helping them to survive at extremely low temperatures. One of the problems in studying such proteins is the difficulty of their isolation and purification. For example, eight cysteine residues in the cfAF (antifreeze protein from the eastern spruce budworm Choristoneura fumiferana) form intermolecular bridges during the overexpression of this protein. This impedes the process of the protein purification dramatically. To overcome this issue, in this work, we designed a mutant form of the ice-binding protein cfAFP, which is much easier to isolate that the wild-type protein. The mutant form named mIBP83 did not lose the ability to bind to ice surface. Besides, observation of the processes of freezing and melting of ice in the presence of mIBP83 showed that this protein affects the process of ice melting, increasing its melting temperature, and does not decrease the water freezing temperature.

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The data presented in this study are available within the article.

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Acknowledgements

We are grateful to Alexei V. Finkelstein for fruitful discussions and Ekaterina N. Samatova for assistance.

Funding

This research was funded by Russian Science Foundation, grant number 21-14-00268.

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Authors and Affiliations

  1. Biophysics Department, Siberian Federal University, Svobodny 79, Krasnoyarsk, Russia, 660041

    Anna A. Deeva & Lala S. Isoyan

  2. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Puschino, Russia

    Ksenia A. Glukhova

  3. Institute of Protein Research of the Russian Academy of Sciences, 4 Institutskaya Str., Pushchino, Moscow Region, Russia, 142290

    Yuliya D. Okulova & Bogdan S. Melnik

  4. Department of Molecular Medicine and Byrd Alzheimer’s Research Institure, Morsani College of Medicine, University of South Florida, Tampa, FL, USA

    Vladimir N. Uversky

Authors
  1. Anna A. Deeva
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  2. Ksenia A. Glukhova
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  3. Lala S. Isoyan
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  4. Yuliya D. Okulova
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  5. Vladimir N. Uversky
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  6. Bogdan S. Melnik
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Contributions

Conceptualization, BSM, VNU; methodology, BSM and VNU; investigation, DAA, ILS, KAG, YDO, BSM, VNU; writing—original draft preparation, BSM and VNU, YDO, DAA, KAG; writing—review and editing, BSM and VNU, YDO, KAG; funding acquisition, BSM All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Bogdan S. Melnik.

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Conflict of interest

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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Deeva, A.A., Glukhova, K.A., Isoyan, L.S. et al. Design and Analysis of a Mutant form of the Ice-Binding Protein from Choristoneura fumiferana. Protein J 41, 304–314 (2022). https://doi.org/10.1007/s10930-022-10049-6

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  • DOI: https://doi.org/10.1007/s10930-022-10049-6

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