Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies

We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purification are use...

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Detalles Bibliográficos
Autores principales: Hakhverdyan, Zhanna, Molloy, Kelly R., Subbotin, Roman I., Fernandez-Martinez, Javier, Chait, Brian T., Rout, Michael P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433251/
https://www.ncbi.nlm.nih.gov/pubmed/34527957
http://dx.doi.org/10.1016/j.xpro.2021.100800
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author Hakhverdyan, Zhanna
Molloy, Kelly R.
Subbotin, Roman I.
Fernandez-Martinez, Javier
Chait, Brian T.
Rout, Michael P.
author_facet Hakhverdyan, Zhanna
Molloy, Kelly R.
Subbotin, Roman I.
Fernandez-Martinez, Javier
Chait, Brian T.
Rout, Michael P.
author_sort Hakhverdyan, Zhanna
collection PubMed
description We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purification are used for the assessment of the whole protein complex dynamics. This method can determine and distinguish both subunit turnover and exchange specifically in an assembly to provide a comprehensive picture of assembly dynamics. For complete details on the use and execution of this protocol, please refer to Hakhverdyan et al. (2021).
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spelling pubmed-84332512021-09-14 Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies Hakhverdyan, Zhanna Molloy, Kelly R. Subbotin, Roman I. Fernandez-Martinez, Javier Chait, Brian T. Rout, Michael P. STAR Protoc Protocol We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purification are used for the assessment of the whole protein complex dynamics. This method can determine and distinguish both subunit turnover and exchange specifically in an assembly to provide a comprehensive picture of assembly dynamics. For complete details on the use and execution of this protocol, please refer to Hakhverdyan et al. (2021). Elsevier 2021-09-08 /pmc/articles/PMC8433251/ /pubmed/34527957 http://dx.doi.org/10.1016/j.xpro.2021.100800 Text en © 2021 The Author(s) 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
Hakhverdyan, Zhanna
Molloy, Kelly R.
Subbotin, Roman I.
Fernandez-Martinez, Javier
Chait, Brian T.
Rout, Michael P.
Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_full Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_fullStr Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_full_unstemmed Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_short Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_sort measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433251/
https://www.ncbi.nlm.nih.gov/pubmed/34527957
http://dx.doi.org/10.1016/j.xpro.2021.100800
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