TY  - JOUR
A1  - D'Imprima, Edoardo
A1  - Floris, Davide
A1  - Joppe, Mirko
A1  - Sánchez, Ricardo
A1  - Grininger, Martin
A1  - Kühlbrandt, Werner
T1  - Protein denaturation at the air-water interface and how to prevent it
T2  - eLife
N2  - 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.
KW  - Research article
KW  - structural biology and molecular biophysics
KW  - cryoem
KW  - graphene
KW  - cryoet
KW  - protein denaturation
KW  - S. Cerevisiae
Y1  - 2019
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/50163
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-501632
SN  - 2050-084X
N1  - 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.
VL  - 8
IS  - e42747
SP  - 1
EP  - 18
PB  - eLife Sciences Publications
CY  - Cambridge
ER  -