Volume 240, 2022

Strategies for picking membrane-associated particles within subtomogram averaging workflows

Abstract

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.

Graphical abstract: Strategies for picking membrane-associated particles within subtomogram averaging workflows

Associated articles

Supplementary files

Article information

Article type
Paper
Submitted
28 Jan 2022
Accepted
04 Mar 2022
First published
12 Apr 2022
This article is Open Access
Creative Commons BY license

Faraday Discuss., 2022,240, 101-113

Strategies for picking membrane-associated particles within subtomogram averaging workflows

E. Pyle, J. Hutchings and G. Zanetti, Faraday Discuss., 2022, 240, 101 DOI: 10.1039/D2FD00022A

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