Abstract
Our understanding of subcellular structures has been greatly increased owing to electron microscopy, even though radiation damage of biological samples by the electron beam demanded staining techniques. Technological and instrumental advances of electron microscopy have, however, established various highly sophisticated techniques to study biological systems in their native states without staining and thus facilitated comprehension of rather intact structures of biological components. Among these techniques, electron crystallography is a well-established one to analyze membrane protein structures within lipid bilayers, without staining at close-to-physiological conditions. Structures of membrane proteins could be analyzed at resolutions better than 3Å by electron crystallography due to techniques of low dose and cryo-electron microscopy (cryo-EM). Here, recent cryo-EM technological and instrumental advances crucial to optimal data collection in electron crystallography are summarized as well as examples of structures of membrane proteins analyzed with the help of this method.
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Acknowledgments
These studies were performed in wonderful collaborations with many researchers whose names were recorded as authors in each referenced paper. This work was supported by Grants-in-Aid for Scientific Research (S) and the Japan New Energy and Industrial Technology Development Organization (NEDO).
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Fujiyoshi, Y. (2013). Low Dose Techniques and Cryo-Electron Microscopy. In: Schmidt-Krey, I., Cheng, Y. (eds) Electron Crystallography of Soluble and Membrane Proteins. Methods in Molecular Biology, vol 955. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-176-9_6
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DOI: https://doi.org/10.1007/978-1-62703-176-9_6
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