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Capturing RNA-dependent pathways for cryo-EM analysis

Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular im...

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Detalles Bibliográficos
Autores principales: Tanner, Justin R., Degen, Katherine, Gilmore, Brian L., Kelly, Deborah F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Research Network of Computational and Structural Biotechnology (RNCSB) Organization 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962177/
https://www.ncbi.nlm.nih.gov/pubmed/24688633
http://dx.doi.org/10.5936/csbj.201204003
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author Tanner, Justin R.
Degen, Katherine
Gilmore, Brian L.
Kelly, Deborah F.
author_facet Tanner, Justin R.
Degen, Katherine
Gilmore, Brian L.
Kelly, Deborah F.
author_sort Tanner, Justin R.
collection PubMed
description Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular imaging platform to capture and view multiple components of cellular pathways within a functionally relevant framework. We utilized the bacterial protein synthesis machinery as a model system to develop our approach. By using modified Affinity Grid surfaces, we were able to recruit multiple protein assemblies bound to nascent strands of mRNA. The combined use of Affinity Capture technology and single particle electron microscopy provide the basis for visualizing RNA-dependent pathways in a remarkable new way.
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spelling pubmed-39621772014-03-31 Capturing RNA-dependent pathways for cryo-EM analysis Tanner, Justin R. Degen, Katherine Gilmore, Brian L. Kelly, Deborah F. Comput Struct Biotechnol J Research Article Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular imaging platform to capture and view multiple components of cellular pathways within a functionally relevant framework. We utilized the bacterial protein synthesis machinery as a model system to develop our approach. By using modified Affinity Grid surfaces, we were able to recruit multiple protein assemblies bound to nascent strands of mRNA. The combined use of Affinity Capture technology and single particle electron microscopy provide the basis for visualizing RNA-dependent pathways in a remarkable new way. Research Network of Computational and Structural Biotechnology (RNCSB) Organization 2012-02-23 /pmc/articles/PMC3962177/ /pubmed/24688633 http://dx.doi.org/10.5936/csbj.201204003 Text en © Tanner et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly cited.
spellingShingle Research Article
Tanner, Justin R.
Degen, Katherine
Gilmore, Brian L.
Kelly, Deborah F.
Capturing RNA-dependent pathways for cryo-EM analysis
title Capturing RNA-dependent pathways for cryo-EM analysis
title_full Capturing RNA-dependent pathways for cryo-EM analysis
title_fullStr Capturing RNA-dependent pathways for cryo-EM analysis
title_full_unstemmed Capturing RNA-dependent pathways for cryo-EM analysis
title_short Capturing RNA-dependent pathways for cryo-EM analysis
title_sort capturing rna-dependent pathways for cryo-em analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962177/
https://www.ncbi.nlm.nih.gov/pubmed/24688633
http://dx.doi.org/10.5936/csbj.201204003
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