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Nat Commun. 2018 Sep 7;9(1):3628. doi: 10.1038/s41467-018-06076-6.

Sub-2 Å Ewald curvature corrected structure of an AAV2 capsid variant.

Author information

1
The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, 89 Convent Ave, New York, NY, 10027, USA.
2
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, 10032, USA.
3
Laboratory of Genetics and Helmsley Center for Genomic Medicine, The Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
4
Department of Biochemistry and Molecular Biology, Center for Structural Biology, The McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, 32610, USA.
5
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
6
Department of Chemistry and Biochemistry and Division of Biological Sciences, University of California-San Diego, La Jolla, CA, 92093, USA.
7
Department of Biochemistry and Molecular Biology, Center for Structural Biology, The McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, 32610, USA. mckenna@ufl.edu.
8
Laboratory of Genetics and Helmsley Center for Genomic Medicine, The Salk Institute for Biological Studies, La Jolla, CA, 92037, USA. dlyumkis@salk.edu.

Abstract

Single-particle cryogenic electron microscopy (cryo-EM) provides a powerful methodology for structural biologists, but the resolutions typically attained with experimentally determined structures have lagged behind microscope capabilities. Here, we exploit several technical advances to improve resolution, including per-particle contrast transfer function (CTF) refinement and correction for Ewald sphere curvature. The latter is demonstrated with several experimental samples and should become more standard as resolutions increase or at lower microscope accelerating voltages. The combined application of the described methods to micrographs recorded on a Titan Krios enables structure determination at ~1.86-Å resolution of an adeno-associated virus serotype 2 variant (AAV2), an important gene-delivery vehicle. The resulting structural details provide an improved model for understanding the biology of AAV that will guide future vector development for gene therapy.

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