Protein denaturation at the air-water interface and how to prevent it
Top Cited Papers
Open Access
- 1 April 2019
- journal article
- research article
- Published by eLife Sciences Publications, Ltd in eLife
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
Funding Information
- Max-Planck-Gesellschaft
- Max-Planck-Gesellschaft
- Volkswagen Foundation
- Alexander von Humboldt Foundation (Sofja Kovalevskaja Award to Mikhail Kudryashev)
- Deutsche Forschungsgemeinschaft (SFB807)
- Volkswagen Foundation (85701)
This publication has 72 references indexed in Scilit:
- Cryomicroscopy of radiation sensitive specimens on unmodified graphene sheets: Reduction of electron-optical effects of chargingJournal of Structural Biology, 2013
- Prevention of overfitting in cryo-EM structure determinationNature Methods, 2012
- The surface of evaporated carbon films is an insulating, high-bandgap materialJournal of Structural Biology, 2011
- Inhibition of the fungal fatty acid synthase type I multienzyme complexProceedings of the National Academy of Sciences of the United States of America, 2008
- Retrospective on the early development of cryoelectron microscopy of macromolecules and a prospective on opportunities for the futureJournal of Structural Biology, 2008
- EMAN2: An extensible image processing suite for electron microscopyJournal of Structural Biology, 2006
- Automated electron microscope tomography using robust prediction of specimen movementsJournal of Structural Biology, 2005
- UCSF Chimera?A visualization system for exploratory research and analysisJournal of Computational Chemistry, 2004
- Computer Visualization of Three-Dimensional Image Data Using IMODJournal of Structural Biology, 1996
- Cryo-electron microscopy of vitrified specimensQuarterly Reviews of Biophysics, 1988