Cargando…

Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM

As the field of electron microscopy advances, the increasing complexity of samples being produced demand more involved processing methods. In this study, we have developed a new processing method for generating 3D reconstructions of tubular structures. Tubular biomolecules are common throughout many...

Descripción completa

Detalles Bibliográficos
Autores principales: Randolph, Peter S., Stagg, Scott M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337063/
https://www.ncbi.nlm.nih.gov/pubmed/32647826
http://dx.doi.org/10.1016/j.yjsbx.2020.100023
_version_ 1783554443303714816
author Randolph, Peter S.
Stagg, Scott M.
author_facet Randolph, Peter S.
Stagg, Scott M.
author_sort Randolph, Peter S.
collection PubMed
description As the field of electron microscopy advances, the increasing complexity of samples being produced demand more involved processing methods. In this study, we have developed a new processing method for generating 3D reconstructions of tubular structures. Tubular biomolecules are common throughout many cellular processes and are appealing targets for biophysical research. Processing of tubules with helical symmetry is relatively straightforward for electron microscopy if the helical parameters are known, but tubular structures that deviate from helical symmetry (asymmetrical components, local but no global order, etc) present myriad issues. Here we present a new processing technique called Reconstruction of Average Subtracted Tubular Regions (RASTR), which was developed to reconstruct tubular structures without applying symmetry. We explain the RASTR approach and quantify its performance using three examples: a simulated symmetrical tubular filament, a symmetrical tubular filament from cryo-EM data, and a membrane tubule coated with locally ordered but not globally ordered proteins.
format Online
Article
Text
id pubmed-7337063
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-73370632020-07-08 Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM Randolph, Peter S. Stagg, Scott M. J Struct Biol X Article As the field of electron microscopy advances, the increasing complexity of samples being produced demand more involved processing methods. In this study, we have developed a new processing method for generating 3D reconstructions of tubular structures. Tubular biomolecules are common throughout many cellular processes and are appealing targets for biophysical research. Processing of tubules with helical symmetry is relatively straightforward for electron microscopy if the helical parameters are known, but tubular structures that deviate from helical symmetry (asymmetrical components, local but no global order, etc) present myriad issues. Here we present a new processing technique called Reconstruction of Average Subtracted Tubular Regions (RASTR), which was developed to reconstruct tubular structures without applying symmetry. We explain the RASTR approach and quantify its performance using three examples: a simulated symmetrical tubular filament, a symmetrical tubular filament from cryo-EM data, and a membrane tubule coated with locally ordered but not globally ordered proteins. Elsevier 2020-03-09 /pmc/articles/PMC7337063/ /pubmed/32647826 http://dx.doi.org/10.1016/j.yjsbx.2020.100023 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Randolph, Peter S.
Stagg, Scott M.
Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title_full Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title_fullStr Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title_full_unstemmed Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title_short Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM
title_sort reconstruction of average subtracted tubular regions (rastr) enables structure determination of tubular filaments by cryo-em
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337063/
https://www.ncbi.nlm.nih.gov/pubmed/32647826
http://dx.doi.org/10.1016/j.yjsbx.2020.100023
work_keys_str_mv AT randolphpeters reconstructionofaveragesubtractedtubularregionsrastrenablesstructuredeterminationoftubularfilamentsbycryoem
AT staggscottm reconstructionofaveragesubtractedtubularregionsrastrenablesstructuredeterminationoftubularfilamentsbycryoem