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Single particle cryo-EM reconstruction of 52 kDa streptavidin at 3.2 Angstrom resolution

By Xiao Fan, Jia Wang, Xing Zhang, Zi Yang, Jin-Can Zhang, Lingyun Zhao, Hai-Lin Peng, Jianlin Lei, Hong-Wei Wang

Posted 13 Nov 2018
bioRxiv DOI: 10.1101/457861 (published DOI: 10.1038/s41467-019-10368-w)

The fast development of single particle cryo-EM has made it more feasible to obtain the 3D structure of well-behaved macromolecules with molecular weight higher than 300 kDa at ~3 Å resolution. It remains a challenge to obtain high resolution structure of molecules smaller than 100 kDa using single particle cryo-EM, mainly due to the low contrast of the molecules embedded in vitreous ice. In this work, we applied the Cs-corrector-VPP coupled cryo-EM to study 52 kDa streptavidin (SA) protein supported on a thin layer of graphene film and embedded in vitreous ice. We were able to solve both the apo-SA and biotin-bound SA at near-atomic resolution using single particle cryo-EM. We demonstrated that the method is capable to determine the structure of molecule as small as 39 kDa and potentially even smaller molecules. Furthermore, we found that using the graphene film to avoid the adsorption to the air-water interface is critical to maintain the protein's high-resolution structural information.

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