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 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.
Metadaten
Author:Edoardo D’Imprima, Davide Floris, Mirko Joppe, Ricardo Sánchez, Martin GriningerORCiDGND, Werner Kühlbrandt
URN:urn:nbn:de:hebis:30:3-501632
DOI:https://doi.org/10.7554/eLife.42747
ISSN:2050-084X
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/30932812
Parent Title (English):eLife
Publisher:eLife Sciences Publications
Place of publication:Cambridge
Contributor(s):Axel T. Brunger
Document Type:Article
Language:English
Year of Completion:2019
Date of first Publication:2019/04/01
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2019/04/18
Tag:Research article; S. Cerevisiae; cryoem; cryoet; graphene; protein denaturation; structural biology and molecular biophysics
Volume:8
Issue:e42747
Page Number:18
First Page:1
Last Page:18
Note:
Copyright D’Imprima et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
HeBIS-PPN:450752208
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Angeschlossene und kooperierende Institutionen / MPI für Biophysik
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 4.0