Microcrystal electron diffraction (MicroED) has recently emerged as a promising method for macromolecular structure determination. The method complements existing structural biology methods by providing the opportunities to study small macromolecular structures (i.e. <50 kDa) from crystals too small for conventional single crystal X-ray diffraction. Several protein structures have been determined and various studies indicate that MicroED is capable of (i) revealing atomic structures with charges, (ii) solving new protein structures by molecular replacement, (iii) visualizing ligand-binding interactions and (iv) solving membrane-protein structures from microcrystals embedded in lipidic mesophases. Since 2019, we have solved two novel protein structures by MicroED[1,3] and shown that it was feasible to use MicroED for structure-based drug discovery[2]. However, comparing to X-ray diffraction, MicroED is still in its infancy. Further optimization and innovations in new software and hardware are required to make MicroED more robust and more accessible to the structural biology community. At Lorne 2023, I will present our latest developments in specimen preparation [4], data collection and processing routine, examples of macromolecular crystal structure determination by MicroED, ongoing method developments and future perspective of MicroED.
[1] H. Xu, et al. Sci. Adv. 5 (2019), eaax4621.
[2] M. T. B. Clabbers, et al. Commun. Biol. 3 (2020), 417.
[3] M. T. B. Clabbers, et al. Nat. Commun. 12 (2021), 2578.
[4] J. Zhao, et al. Nat. Commun. 12 (2021), 5036.