Protein denaturation at the air-water interface and how to prevent it.
| Autor: | D'Imprima E; Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, Germany., Floris D; Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, Germany., Joppe M; Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt, Germany., Sánchez R; Sofja Kovalevskaja Group, Max Planck Institute of Biophysics, Frankfurt, Germany., Grininger M; Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt, Germany., Kühlbrandt W; Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, Germany. |
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| Jazyk: | angličtina |
| Zdroj: | ELife [Elife] 2019 Apr 01; Vol. 8. Date of Electronic Publication: 2019 Apr 01. |
| DOI: | 10.7554/eLife.42747 |
| Abstrakt: | 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. Competing Interests: ED, DF, MJ, RS, MG No competing interests declared, WK Reviewing editor, eLife (© 2019, D'Imprima et al.) |
| Databáze: | MEDLINE |
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