An integrated workflow for charting the human interaction proteome: insights into the PP2A system

Abstract Protein complexes represent major functional units for the execution of biological processes. Systematic affinity purification coupled with mass spectrometry (AP‐MS) yielded a wealth of information on the compendium of protein complexes expressed in Saccharomyces cerevisiae. However, global...

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Main Authors: Timo Glatter, Alexander Wepf, Ruedi Aebersold, Matthias Gstaiger
Format: Article
Language:English
Published: Springer Nature 2009-01-01
Series:Molecular Systems Biology
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Online Access:https://doi.org/10.1038/msb.2008.75
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author Timo Glatter
Alexander Wepf
Ruedi Aebersold
Matthias Gstaiger
author_facet Timo Glatter
Alexander Wepf
Ruedi Aebersold
Matthias Gstaiger
author_sort Timo Glatter
collection DOAJ
description Abstract Protein complexes represent major functional units for the execution of biological processes. Systematic affinity purification coupled with mass spectrometry (AP‐MS) yielded a wealth of information on the compendium of protein complexes expressed in Saccharomyces cerevisiae. However, global AP‐MS analysis of human protein complexes is hampered by the low throughput, sensitivity and data robustness of existing procedures, which limit its application for systems biology research. Here, we address these limitations by a novel integrated method, which we applied and benchmarked for the human protein phosphatase 2A system. We identified a total of 197 protein interactions with high reproducibility, showing the coexistence of distinct classes of phosphatase complexes that are linked to proteins implicated in mitosis, cell signalling, DNA damage control and more. These results show that the presented analytical process will substantially advance throughput and reproducibility in future systematic AP‐MS studies on human protein complexes.
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spelling doaj-art-0bdef0c2f00342f8ade8e4e8a7705cf72025-08-24T11:58:58ZengSpringer NatureMolecular Systems Biology1744-42922009-01-015111410.1038/msb.2008.75An integrated workflow for charting the human interaction proteome: insights into the PP2A systemTimo Glatter0Alexander Wepf1Ruedi Aebersold2Matthias Gstaiger3Institute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichAbstract Protein complexes represent major functional units for the execution of biological processes. Systematic affinity purification coupled with mass spectrometry (AP‐MS) yielded a wealth of information on the compendium of protein complexes expressed in Saccharomyces cerevisiae. However, global AP‐MS analysis of human protein complexes is hampered by the low throughput, sensitivity and data robustness of existing procedures, which limit its application for systems biology research. Here, we address these limitations by a novel integrated method, which we applied and benchmarked for the human protein phosphatase 2A system. We identified a total of 197 protein interactions with high reproducibility, showing the coexistence of distinct classes of phosphatase complexes that are linked to proteins implicated in mitosis, cell signalling, DNA damage control and more. These results show that the presented analytical process will substantially advance throughput and reproducibility in future systematic AP‐MS studies on human protein complexes.https://doi.org/10.1038/msb.2008.75human interactomemass spectrometryPP2Aprotein complexes
spellingShingle Timo Glatter
Alexander Wepf
Ruedi Aebersold
Matthias Gstaiger
An integrated workflow for charting the human interaction proteome: insights into the PP2A system
Molecular Systems Biology
human interactome
mass spectrometry
PP2A
protein complexes
title An integrated workflow for charting the human interaction proteome: insights into the PP2A system
title_full An integrated workflow for charting the human interaction proteome: insights into the PP2A system
title_fullStr An integrated workflow for charting the human interaction proteome: insights into the PP2A system
title_full_unstemmed An integrated workflow for charting the human interaction proteome: insights into the PP2A system
title_short An integrated workflow for charting the human interaction proteome: insights into the PP2A system
title_sort integrated workflow for charting the human interaction proteome insights into the pp2a system
topic human interactome
mass spectrometry
PP2A
protein complexes
url https://doi.org/10.1038/msb.2008.75
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