The interior of eukaryotic cells is mysterious. How do the large communities of macromolecular machines interact with each other? How do the structures and positions of these nanoscopic entities respond to new stimuli? Questions like these can now be answered with the help of a method called electron cryotomography (cryo-ET). Cryo-ET will ultimately reveal the inner workings of a cell at the protein, secondary structure, and perhaps even side-chain levels. Combined with genetic or pharmacological perturbation, cryo-ET will allow us to answer previously unimaginable questions, such as how structure, biochemistry, and forces are related in situ. Because it bridges structural biology and cell biology, cryo-ET is indispensable for structural cell biology—the study of the 3-D macromolecular structure of cells. Here we discuss some of the key ideas, strategies, auxiliary techniques, and innovations that an aspiring structural cell biologist will consider when planning to ask bold questions.
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Downloads: 523History Submitted: 12 February 2019Revised: 25 November 2019Accepted: 29 November 2019
Information © 2020 Ng and Gan. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
We thank Eunyoung Chae, Min Wu, Tony Kanchanawong, Shu Fen Tan, and Alasdair McDowall for discussions and the anonymous reviewers for their constructive feedback. The images and movies were created with Bsoft, IMOD, UCSF Chimera, avconv, and Adobe Creative Cloud ( Mastronarde, 1997; Pettersen et al., 2004; Heymann and Belnap, 2007; Libav_team, 2018). All panels in Figures 3– 5 were adapted with permission from the authors and publishers. This work was supported by Singapore Ministry of Education T1 R-154-000-A49-114 and MOE2018-T2-2-146.
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