Mass-selective and ice-free electron cryomicroscopy protein sample preparation via native electrospray ion-beam deposition.

ABSTRACT: Despite tremendous advances in sample preparation and classification algorithms for electron cryomicroscopy (cryo-EM) and single-particle analysis (SPA), sample heterogeneity remains a major challenge and can prevent access to high-resolution structures. In addition, optimization of preparation conditions for a given sample can be time-consuming. In the current work, it is demonstrated that native electrospray ion-beam deposition (native ES-IBD) is an alternative, reliable approach for the preparation of extremely high-purity samples, based on mass selection in vacuum. Folded protein ions are generated by native electrospray ionization, separated from other proteins, contaminants, aggregates, and fragments, gently deposited on cryo-EM grids, frozen in liquid nitrogen, and subsequently imag

SUBMITTER: Esser TK 

PROVIDER: S-EPMC9802471 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature