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...

Full description

Bibliographic Details
Main Authors:	Edoardo D'Imprima, Davide Floris, Mirko Joppe, Ricardo Sánchez, Martin Grininger, Werner Kühlbrandt
Format:	Article
Language:	English
Published:	eLife Sciences Publications Ltd 2019-04-01
Series:	eLife
Subjects:	CryoEM Graphene CryoET Protein denaturation
Online Access:	https://elifesciences.org/articles/42747

id	doaj-b75524f2a8de4dbab311677c9269b551
record_format	Article
spelling	doaj-b75524f2a8de4dbab311677c9269b5512021-05-05T17:30:22ZengeLife 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
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Edoardo D'Imprima Davide Floris Mirko Joppe Ricardo Sánchez Martin Grininger Werner Kühlbrandt
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
author_facet	Edoardo D'Imprima Davide Floris Mirko Joppe Ricardo Sánchez Martin Grininger Werner Kühlbrandt
author_sort	Edoardo D'Imprima
title	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_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_sort	protein denaturation at the air-water interface and how to prevent it
publisher	eLife Sciences Publications Ltd
series	eLife
issn	2050-084X
publishDate	2019-04-01
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.
topic	CryoEM Graphene CryoET Protein denaturation
url	https://elifesciences.org/articles/42747
work_keys_str_mv	AT edoardodimprima proteindenaturationattheairwaterinterfaceandhowtopreventit AT davidefloris proteindenaturationattheairwaterinterfaceandhowtopreventit AT mirkojoppe proteindenaturationattheairwaterinterfaceandhowtopreventit AT ricardosanchez proteindenaturationattheairwaterinterfaceandhowtopreventit AT martingrininger proteindenaturationattheairwaterinterfaceandhowtopreventit AT wernerkuhlbrandt proteindenaturationattheairwaterinterfaceandhowtopreventit
_version_	1721459237540855808

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

Similar Items