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Direct evidence for pinning of single, ice-bound antifreeze proteins by subzero nanoscopy

View ORCID ProfileRoderick P. Tas, Marco M. R. M. Hendrix, View ORCID ProfileIlja K. Voets
doi: https://doi.org/10.1101/2022.04.05.487137
Roderick P. Tas
1Laboratory of Self-Organizing Soft Matter, Department of Chemical Engineering and Chemistry, and Institute for Complex Molecular Systems, Eindhoven University of Technology (TUE), Eindhoven, 5612 AP, The Netherlands
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  • For correspondence: r.p.tas@tue.nl i.voets@tue.nl
Marco M. R. M. Hendrix
1Laboratory of Self-Organizing Soft Matter, Department of Chemical Engineering and Chemistry, and Institute for Complex Molecular Systems, Eindhoven University of Technology (TUE), Eindhoven, 5612 AP, The Netherlands
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Ilja K. Voets
1Laboratory of Self-Organizing Soft Matter, Department of Chemical Engineering and Chemistry, and Institute for Complex Molecular Systems, Eindhoven University of Technology (TUE), Eindhoven, 5612 AP, The Netherlands
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  • ORCID record for Ilja K. Voets
  • For correspondence: r.p.tas@tue.nl i.voets@tue.nl
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Abstract

Ice-binding by antifreeze proteins (AFPs) reduces freezing temperatures and arrests ice-crystal ripening, making AFPs essential for survival in ice-laden environments and attractive as biocompatible antifreezes. Whilst their activity was identified over 50 years ago, the physical mechanisms are still debated because experimental insights at the molecular scale remain elusive. Here we introduce optical nanoscopy to resolve the ice/water interfacial dynamics of single AFPs. Using this method, we demonstrate pinning of individual proteins. Surprisingly, this quasi-permanent pinning is lost when freezing point depression activity is inhibited by a single mutation in the ice-binding site. These findings provide direct experimental evidence for the adsorption-inhibition paradigm, pivotal to all theoretical descriptions of activity and offer new insights in the molecular mechanisms by which these biological antifreezes function.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 07, 2022.
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Direct evidence for pinning of single, ice-bound antifreeze proteins by subzero nanoscopy
Roderick P. Tas, Marco M. R. M. Hendrix, Ilja K. Voets
bioRxiv 2022.04.05.487137; doi: https://doi.org/10.1101/2022.04.05.487137
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Direct evidence for pinning of single, ice-bound antifreeze proteins by subzero nanoscopy
Roderick P. Tas, Marco M. R. M. Hendrix, Ilja K. Voets
bioRxiv 2022.04.05.487137; doi: https://doi.org/10.1101/2022.04.05.487137

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