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Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM.
Sanchez, Ricardo M; Zhang, Yingyi; Chen, Wenbo; Dietrich, Lea; Kudryashev, Mikhail.
Afiliação
  • Sanchez RM; Max Planck Institute for Biophysics, Max-von-Laue Strasse, 3, 60348, Frankfurt am Main, Germany.
  • Zhang Y; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt am Main, Max-von-Laue Strasse, 15, 60348, Frankfurt am Main, Germany.
  • Chen W; Max Planck Institute for Biophysics, Max-von-Laue Strasse, 3, 60348, Frankfurt am Main, Germany.
  • Dietrich L; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt am Main, Max-von-Laue Strasse, 15, 60348, Frankfurt am Main, Germany.
  • Kudryashev M; Max Planck Institute for Biophysics, Max-von-Laue Strasse, 3, 60348, Frankfurt am Main, Germany.
Nat Commun ; 11(1): 3709, 2020 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-32709843
ABSTRACT
Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace due to beam-induced sample drift, images with poor signal-to-noise ratios (SNR), challenges in CTF correction, and limited particle number. Here we address these issues by collecting tilt series with a higher electron dose at the zero-degree tilt. Particles of interest are then located within reconstructed tomograms, processed by conventional StA, and then re-extracted from the high-dose images in 2D. Single particle analysis tools are then applied to refine the 2D particle alignment and generate a reconstruction. Use of our hybrid StA (hStA) workflow improved the resolution for tobacco mosaic virus from 7.2 to 4.4 Å and for the ion channel RyR1 in crowded native membranes from 12.9 to 9.1 Å. These resolution gains make hStA a promising approach for other StA projects aimed at achieving subnanometer resolution.
Assuntos
Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: Processamento de Imagem Assistida por Computador / Canais de Cálcio / Microscopia Crioeletrônica Idioma: Inglês Revista: Nat Commun Assunto da revista: Biologia / Ciência Ano de publicação: 2020 Tipo de documento: Artigo País de afiliação: Alemanha

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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: Processamento de Imagem Assistida por Computador / Canais de Cálcio / Microscopia Crioeletrônica Idioma: Inglês Revista: Nat Commun Assunto da revista: Biologia / Ciência Ano de publicação: 2020 Tipo de documento: Artigo País de afiliação: Alemanha
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