Open architecture of archaea MCM and dsDNA complexes resolved using monodispersed streptavidin affinity CryoEM.

ABSTRACT: The cryo-electron microscopy (cryoEM) method has enabled high-resolution structure determination of numerous biomolecules and complexes. Nevertheless, cryoEM sample preparation of challenging proteins and complexes, especially those with low abundance or with preferential orientation, remains a major hurdle. We developed an affinity-grid method employing monodispersed single particle streptavidin on a lipid monolayer to enhance particle absorption on the grid surface and alleviate sample exposure to the air-water interface. Using this approach, we successfully enriched the Thermococcus kodakarensis mini-chromosome maintenance complex 3 (MCM3) on cryoEM grids through biotinylation and resolved its structure. We further utilized this affinity method to tether the biotin-tagged dsDNA to selec

SUBMITTER: Ma J 

PROVIDER: S-EPMC11603195 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature