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TitleProtein denaturation at the air-water interface and how to prevent it.
Journal, issue, pagesElife, Vol. 8, Year 2019
Publish dateApr 1, 2019
AuthorsEdoardo D'Imprima / Davide Floris / Mirko Joppe / Ricardo Sánchez / Martin Grininger / Werner Kühlbrandt /
PubMed AbstractElectron 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, ...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.
External linksElife / PubMed:30932812 / PubMed Central
MethodsEM (single particle) / EM (subtomogram averaging)
Resolution4.0 - 24.6 Å
Structure data

EMDB-0178:
Cryo-EM map of the Fatty Acid Synthase from S. cerevisiae
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-0179:
Cryo-EM map of the Fatty Acid Synthase from S. cerevisiae
Method: EM (subtomogram averaging) / Resolution: 24.6 Å

Source
  • Saccharomyces cerevisiae (brewer's yeast)

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