Using a partial atomic model from medium-resolution cryo-EM to solve a large crystal structure

Acta Crystallogr D Struct Biol. 2021 Jan 1;77(Pt 1):11-18. doi: 10.1107/S2059798320015156. Epub 2021 Jan 1.

Abstract

Medium-resolution cryo-electron microscopy maps, in particular when they include a significant number of α-helices, may allow the building of partial models that are useful for molecular-replacement searches in large crystallographic structures when the structures of homologs are not available and experimental phasing has failed. Here, as an example, the solution of the structure of a bacteriophage portal using a partial 30% model built into a 7.8 Å resolution cryo-EM map is shown. Inspection of the self-rotation function allowed the correct oligomerization state to be determined, and density-modification procedures using rotation matrices and a mask based on the cryo-EM structure were critical for solving the structure. A workflow is described that may be applicable to similar cases and this strategy is compared with direct use of the cryo-EM map for molecular replacement.

Keywords: bacteriophage portal; cryo-EM; density modification; molecular replacement.

MeSH terms

  • Bacteriophage T7 / metabolism*
  • Capsid Proteins / chemistry*
  • Cryoelectron Microscopy / methods*
  • Models, Molecular
  • Protein Conformation
  • Software

Substances

  • Capsid Proteins