Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast

Here, we describe a fractionation protocol optimized to quantify changes in relative abundance of the chromatin-bound proteome (chromatome) by tandem mass tag multiplexing-based tandem mass spectrometry. It has been applied to yeast cells before and after exposure to DNA-damaging drugs to characteri...

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Detalles Bibliográficos
Autores principales: Challa, Kiran, Seebacher, Jan, Gasser, Susan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449121/
https://www.ncbi.nlm.nih.gov/pubmed/34568845
http://dx.doi.org/10.1016/j.xpro.2021.100825
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author Challa, Kiran
Seebacher, Jan
Gasser, Susan M.
author_facet Challa, Kiran
Seebacher, Jan
Gasser, Susan M.
author_sort Challa, Kiran
collection PubMed
description Here, we describe a fractionation protocol optimized to quantify changes in relative abundance of the chromatin-bound proteome (chromatome) by tandem mass tag multiplexing-based tandem mass spectrometry. It has been applied to yeast cells before and after exposure to DNA-damaging drugs to characterize changes in chromatin composition induced by the DNA damage response. We detail steps for stringent chromatin fractionation, sample preparation for mass spectrometry, and its evaluation. For complete details on the use and execution of this protocol, please refer to Challa et al. (2021).
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spelling pubmed-84491212021-09-24 Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast Challa, Kiran Seebacher, Jan Gasser, Susan M. STAR Protoc Protocol Here, we describe a fractionation protocol optimized to quantify changes in relative abundance of the chromatin-bound proteome (chromatome) by tandem mass tag multiplexing-based tandem mass spectrometry. It has been applied to yeast cells before and after exposure to DNA-damaging drugs to characterize changes in chromatin composition induced by the DNA damage response. We detail steps for stringent chromatin fractionation, sample preparation for mass spectrometry, and its evaluation. For complete details on the use and execution of this protocol, please refer to Challa et al. (2021). Elsevier 2021-09-14 /pmc/articles/PMC8449121/ /pubmed/34568845 http://dx.doi.org/10.1016/j.xpro.2021.100825 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
Challa, Kiran
Seebacher, Jan
Gasser, Susan M.
Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title_full Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title_fullStr Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title_full_unstemmed Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title_short Sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
title_sort sucrose gradient chromatin enrichment for quantitative proteomics analysis in budding yeast
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449121/
https://www.ncbi.nlm.nih.gov/pubmed/34568845
http://dx.doi.org/10.1016/j.xpro.2021.100825
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