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Elife. 2019 Apr 1;8. pii: e42747. doi: 10.7554/eLife.42747.

Protein denaturation at the air-water interface and how to prevent it.

Author information

1
Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, Germany.
2
Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt, Germany.
3
Sofja Kovalevskaja Group, Max Planck Institute of Biophysics, Frankfurt, Germany.
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Contributed equally

Abstract

Electron cryo-microscopy analyzes the structure of proteins and protein complexes in vitrified solution. Proteins tend to adsorb to the air-water interface in unsupported films of aqueous solution, which can result in partial or complete denaturation. We investigated the structure of yeast fatty acid synthase at the air-water interface by electron cryo-tomography and single-particle image processing. Around 90% of complexes adsorbed to the air-water interface are partly denatured. We show that the unfolded regions face the air-water interface. Denaturation by contact with air may happen at any stage of specimen preparation. Denaturation at the air-water interface is completely avoided when the complex is plunge-frozen on a substrate of hydrophilized graphene.

KEYWORDS:

CryoEM; CryoET; Graphene; Protein denaturation; S. cerevisiae; molecular biophysics; structural biology

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