Protein denaturation at the air-water interface and how to prevent it
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 aci...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-04-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/42747 |
| _version_ | 1811181046615506944 |
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| author | Edoardo D'Imprima Davide Floris Mirko Joppe Ricardo Sánchez Martin Grininger Werner Kühlbrandt |
| author_facet | Edoardo D'Imprima Davide Floris Mirko Joppe Ricardo Sánchez Martin Grininger Werner Kühlbrandt |
| author_sort | Edoardo D'Imprima |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:13:02Z |
| format | Article |
| id | doaj.art-b75524f2a8de4dbab311677c9269b551 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:13:02Z |
| publishDate | 2019-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-b75524f2a8de4dbab311677c9269b5512022-12-22T04:32:27ZengeLife Sciences Publications LtdeLife2050-084X2019-04-01810.7554/eLife.42747Protein denaturation at the air-water interface and how to prevent itEdoardo D'Imprima0https://orcid.org/0000-0002-9830-7929Davide Floris1https://orcid.org/0000-0001-9144-5459Mirko Joppe2https://orcid.org/0000-0001-6463-0253Ricardo Sánchez3Martin Grininger4Werner Kühlbrandt5https://orcid.org/0000-0002-2013-4810Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, GermanyDepartment of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, GermanyBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt, GermanySofja Kovalevskaja Group, Max Planck Institute of Biophysics, Frankfurt, GermanyBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt, GermanyDepartment of Structural Biology, Max Planck Institute of Biophysics, Frankfurt, GermanyElectron 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.https://elifesciences.org/articles/42747CryoEMGrapheneCryoETProtein denaturation |
| spellingShingle | Edoardo D'Imprima Davide Floris Mirko Joppe Ricardo Sánchez Martin Grininger Werner Kühlbrandt Protein denaturation at the air-water interface and how to prevent it eLife CryoEM Graphene CryoET Protein denaturation |
| title | Protein denaturation at the air-water interface and how to prevent it |
| title_full | Protein denaturation at the air-water interface and how to prevent it |
| title_fullStr | Protein denaturation at the air-water interface and how to prevent it |
| title_full_unstemmed | Protein denaturation at the air-water interface and how to prevent it |
| title_short | Protein denaturation at the air-water interface and how to prevent it |
| title_sort | protein denaturation at the air water interface and how to prevent it |
| topic | CryoEM Graphene CryoET Protein denaturation |
| url | https://elifesciences.org/articles/42747 |
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