A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells

The functional and structural characterization of macromolecular complexes requires protocols for their native isolation. Here, we describe a protocol for this task based on the recombinant poxvirus Vaccinia expressing tagged proteins of interest in infected cells. Tagged proteins and their interact...

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Detalles Bibliográficos
Autores principales: Bartuli, Julia, Lorenzi, Isotta, Backes, Simone, Grimm, Clemens, Fischer, Utz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792428/
https://www.ncbi.nlm.nih.gov/pubmed/35118428
http://dx.doi.org/10.1016/j.xpro.2021.101116
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author Bartuli, Julia
Lorenzi, Isotta
Backes, Simone
Grimm, Clemens
Fischer, Utz
author_facet Bartuli, Julia
Lorenzi, Isotta
Backes, Simone
Grimm, Clemens
Fischer, Utz
author_sort Bartuli, Julia
collection PubMed
description The functional and structural characterization of macromolecular complexes requires protocols for their native isolation. Here, we describe a protocol for this task based on the recombinant poxvirus Vaccinia expressing tagged proteins of interest in infected cells. Tagged proteins and their interactors can then be isolated via affinity chromatography. The procedure is illustrated for the Vaccinia virus encoded multi-subunit RNA polymerase. Our protocol also allows the expression and isolation of heterologous proteins and hence is suitable for a broader application. For complete details on the use and execution of this profile, please refer to Grimm et al. (2019).
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spelling pubmed-87924282022-02-02 A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells Bartuli, Julia Lorenzi, Isotta Backes, Simone Grimm, Clemens Fischer, Utz STAR Protoc Protocol The functional and structural characterization of macromolecular complexes requires protocols for their native isolation. Here, we describe a protocol for this task based on the recombinant poxvirus Vaccinia expressing tagged proteins of interest in infected cells. Tagged proteins and their interactors can then be isolated via affinity chromatography. The procedure is illustrated for the Vaccinia virus encoded multi-subunit RNA polymerase. Our protocol also allows the expression and isolation of heterologous proteins and hence is suitable for a broader application. For complete details on the use and execution of this profile, please refer to Grimm et al. (2019). Elsevier 2022-01-21 /pmc/articles/PMC8792428/ /pubmed/35118428 http://dx.doi.org/10.1016/j.xpro.2021.101116 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Bartuli, Julia
Lorenzi, Isotta
Backes, Simone
Grimm, Clemens
Fischer, Utz
A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title_full A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title_fullStr A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title_full_unstemmed A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title_short A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
title_sort generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792428/
https://www.ncbi.nlm.nih.gov/pubmed/35118428
http://dx.doi.org/10.1016/j.xpro.2021.101116
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