Kavli Affiliate: Carol Robinson
| Summary:
Electrospray ion beam deposition (ESIBD) is the intact, chemically selective deposition of molecular ions on surfaces in vacuum. Here, we present a general method and dedicated instrumentation for ESIBD-based cryoEM sample preparation of soluble proteins. Precise control over deposition energy, sample environment, and reproducible growth of thin, homogeneous, vitreous ice films embedding the deposited proteins results in samples suitable for high-resolution cryoEM structure determination. Applied to several protein complexes, $β$-Galactosidase, GDH, RuBisCo, GroEL, the workflow yields near-atomic resolution cryoEM maps (2.5-4.8$,Å$) from which atomic models are derived. Dehydration-induced structural changes correlate with the magnitude of solvent exposure in the native structure: interior residues present high-resolution density while surface-exposed regions rearrange. Coherent rearrangements retain secondary and tertiary structure, incoherent changes degrade resolution. These results establish ESIBD+cryoEM as viable method for structure determination of chemically selected protein samples, directly linking native MS chemical information with near-atomic structural resolution.
| Search Query: arXiv Query: search_query=au:”Robinson Carol”&id_list=&start=0&max_results=10
Read More