The deadly touch: protein denaturation at the air-water interface and how to prevent it

Datum

21.01.2019

Zeit

11:00 - 12:00

Sprecher

Edoardo D’Imprima

Zugehörigkeit

Structural Biology Department, Max Planck Institute of Biophysics, Frankfurt

Sprache

en

Hauptthema

Biologie

Andere Themen

Biologie

Host

Gaia Pigino

Beschreibung

Electron cryo-microscopy analyzes the structure of proteins and protein complexes in vitrified solution. Proteins tend to be adsorbed to the air-water interface in unsupported film of aqueous solution, which can result in partial or complete denaturation of the macromolecular complex. We investigated the structure of Saccharomyces cerevisiae fatty acid synthase (FAS) 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. Combining our results in cryo-conditions with negative stain experiments we demonstrate that 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.

Letztmalig verändert: 22.01.2019, 01:08:41