It started with a Cys: Spontaneous cysteine modification during cryo-EM grid preparation.

ABSTRACT: Advances in single particle cryo-EM data collection and processing have seen a significant rise in its use. However, the influences of the environment generated through grid preparation, by for example interactions of proteins with the air-water interface are poorly understood and can be a major hurdle in structure determination by cryo-EM. Initial interactions of proteins with the air-water interface occur quickly and proteins can adopt preferred orientation or partially unfold within hundreds of milliseconds. It has also been shown previously that thin-film layers create hydroxyl radicals. To investigate the potential this might have in cryo-EM sample preparation, we studied two proteins, HSPD1, and beta-galactosidase, and show that cysteine residues are modified in a time-dependent mann

SUBMITTER: Klebl DP 

PROVIDER: S-EPMC9389043 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature