4.2 Article

Strategies for picking membrane-associated particles within subtomogram averaging workflows

期刊

FARADAY DISCUSSIONS
卷 240, 期 -, 页码 101-113

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ROYAL SOC CHEMISTRY
DOI: 10.1039/d2fd00022a

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资金

  1. ERC [StG 852915-CRYTOCOP]
  2. BBSRC [BB/T002670/1]

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This article presents a time-efficient and accurate strategy for picking and cleaning membrane-associated particles and provides a step-by-step guide and necessary scripts for users to carry out these strategies.
Cryo-electron tomography (cryo-ET) with subtomogram averaging (STA) has emerged as a key tool for determining macromolecular structure(s) in vitro and in situ. However, processing cryo-ET data with STA currently requires significant user expertise. Recent efforts have streamlined several steps in STA workflows; however, particle picking remains a time-consuming bottleneck for many projects and requires considerable user input. Here, we present several strategies for the time-efficient and accurate picking of membrane-associated particles using the COPII inner coat as a case study. We also discuss a range of particle cleaning solutions to remove both poor quality and false-positive particles from STA datasets. We provide a step-by-step guide and the necessary scripts for users to independently carry out the particle picking and cleaning strategies discussed.

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