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Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions amo...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8808283/ https://www.ncbi.nlm.nih.gov/pubmed/35128479 http://dx.doi.org/10.1016/j.xpro.2022.101137 |
_version_ | 1784643854903803904 |
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author | Cipak, Lubos Selicky, Tomas Jurcik, Jan Cipakova, Ingrid Osadska, Michaela Lukacova, Veronika Barath, Peter Gregan, Juraj |
author_facet | Cipak, Lubos Selicky, Tomas Jurcik, Jan Cipakova, Ingrid Osadska, Michaela Lukacova, Veronika Barath, Peter Gregan, Juraj |
author_sort | Cipak, Lubos |
collection | PubMed |
description | Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions among proteins. Here, we present a protocol for tandem affinity purification (TAP) of protein complexes from the fission yeast Schizosaccharomyces pombe. The protocol employs cells expressing C-terminally TAP-tagged proteins and is suitable for the analysis of purified proteins by mass spectrometry. For complete information on the use and execution of this protocol, please refer to Cipakova et al. (2019). |
format | Online Article Text |
id | pubmed-8808283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-88082832022-02-04 Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe Cipak, Lubos Selicky, Tomas Jurcik, Jan Cipakova, Ingrid Osadska, Michaela Lukacova, Veronika Barath, Peter Gregan, Juraj STAR Protoc Protocol Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions among proteins. Here, we present a protocol for tandem affinity purification (TAP) of protein complexes from the fission yeast Schizosaccharomyces pombe. The protocol employs cells expressing C-terminally TAP-tagged proteins and is suitable for the analysis of purified proteins by mass spectrometry. For complete information on the use and execution of this protocol, please refer to Cipakova et al. (2019). Elsevier 2022-01-28 /pmc/articles/PMC8808283/ /pubmed/35128479 http://dx.doi.org/10.1016/j.xpro.2022.101137 Text en © 2022 The Authors https://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 | Protocol Cipak, Lubos Selicky, Tomas Jurcik, Jan Cipakova, Ingrid Osadska, Michaela Lukacova, Veronika Barath, Peter Gregan, Juraj Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title | Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title_full | Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title_fullStr | Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title_full_unstemmed | Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title_short | Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe |
title_sort | tandem affinity purification protocol for isolation of protein complexes from schizosaccharomyces pombe |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8808283/ https://www.ncbi.nlm.nih.gov/pubmed/35128479 http://dx.doi.org/10.1016/j.xpro.2022.101137 |
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