Journal
NATURE
Volume 587, Issue 7832, Pages 152-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41586-020-2829-0
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Funding
- EM facilities at the MRC-LMB
- Biochemistry Department of Cambridge University
- Thermo Fisher Scientific
- Wellcome Trust [206171/Z/17/Z, 202905/Z/16/Z]
- University of Cambridge
- UK Medical Research Council [MC_UP_A025_1012, MR/L009609/1, MC_UP_1201/15, MC_UP_A025_1013]
- Japan Society for the Promotion of Science (Overseas Research Fellowship)
- MRC-LMB
- Cambridge Trust
- School of Clinical Medicine, University of Cambridge (LMB Cambridge Scholarship)
- School of Clinical Medicine, University of Cambridge (Cambridge MB/PhD fellowship)
- European Commission (Marie Skodowska-Curie Actions) [H2020-MSCA-IF-2015/709054, H2020-MSCA-IF-2017/793653]
- EMBO [300-2015]
- Cancer Research UK [C20724/A14414, C20724/A26752]
- Boehringer Ingelheim Fonds
- Research Foundation - Flanders (FWO)
- BBSRC [BB/M024709/1] Funding Source: UKRI
- MRC [MC_UP_1201/15, MC_UP_A025_1012, MR/L009609/1, MC_UP_A025_1013] Funding Source: UKRI
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The three-dimensional positions of atoms in protein molecules define their structure and their roles in biological processes. The more precisely atomic coordinates are determined, the more chemical information can be derived and the more mechanistic insights into protein function may be inferred. Electron cryo-microscopy (cryo-EM) single-particle analysis has yielded protein structures with increasing levels of detail in recent years(1,2). However, it has proved difficult to obtain cryo-EM reconstructions with sufficient resolution to visualize individual atoms in proteins. Here we use a new electron source, energy filter and camera to obtain a 1.7 angstrom resolution cryo-EM reconstruction for a human membrane protein, the beta 3 GABA(A)receptor homopentamer(3). Such maps allow a detailed understanding of small-molecule coordination, visualization of solvent molecules and alternative conformations for multiple amino acids, and unambiguous building of ordered acidic side chains and glycans. Applied to mouse apoferritin, our strategy led to a 1.22 angstrom resolution reconstruction that offers a genuine atomic-resolution view of a protein molecule using single-particle cryo-EM. Moreover, the scattering potential from many hydrogen atoms can be visualized in difference maps, allowing a direct analysis of hydrogen-bonding networks. Our technological advances, combined with further approaches to accelerate data acquisition and improve sample quality, provide a route towards routine application of cryo-EM in high-throughput screening of small molecule modulators and structure-based drug discovery. Advances in electron cryo-microscopy hardware allow proteins to be studied at atomic resolution.
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