Capturing protein communities by structural proteomics in a thermophilic eukaryote

Abstract The arrangement of proteins into complexes is a key organizational principle for many cellular functions. Although the topology of many complexes has been systematically analyzed in isolation, their molecular sociology in situ remains elusive. Here, we show that crude cellular extracts of a...

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Main Authors: Panagiotis L Kastritis, Francis J O'Reilly, Thomas Bock, Yuanyue Li, Matt Z Rogon, Katarzyna Buczak, Natalie Romanov, Matthew J Betts, Khanh Huy Bui, Wim J Hagen, Marco L Hennrich, Marie‐Therese Mackmull, Juri Rappsilber, Robert B Russell, Peer Bork, Martin Beck, Anne‐Claude Gavin
Format: Article
Language:English
Published: Springer Nature 2017-07-01
Series:Molecular Systems Biology
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Online Access:https://doi.org/10.15252/msb.20167412
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author Panagiotis L Kastritis
Francis J O'Reilly
Thomas Bock
Yuanyue Li
Matt Z Rogon
Katarzyna Buczak
Natalie Romanov
Matthew J Betts
Khanh Huy Bui
Wim J Hagen
Marco L Hennrich
Marie‐Therese Mackmull
Juri Rappsilber
Robert B Russell
Peer Bork
Martin Beck
Anne‐Claude Gavin
author_facet Panagiotis L Kastritis
Francis J O'Reilly
Thomas Bock
Yuanyue Li
Matt Z Rogon
Katarzyna Buczak
Natalie Romanov
Matthew J Betts
Khanh Huy Bui
Wim J Hagen
Marco L Hennrich
Marie‐Therese Mackmull
Juri Rappsilber
Robert B Russell
Peer Bork
Martin Beck
Anne‐Claude Gavin
author_sort Panagiotis L Kastritis
collection DOAJ
description Abstract The arrangement of proteins into complexes is a key organizational principle for many cellular functions. Although the topology of many complexes has been systematically analyzed in isolation, their molecular sociology in situ remains elusive. Here, we show that crude cellular extracts of a eukaryotic thermophile, Chaetomium thermophilum, retain basic principles of cellular organization. Using a structural proteomics approach, we simultaneously characterized the abundance, interactions, and structure of a third of the C. thermophilum proteome within these extracts. We identified 27 distinct protein communities that include 108 interconnected complexes, which dynamically associate with each other and functionally benefit from being in close proximity in the cell. Furthermore, we investigated the structure of fatty acid synthase within these extracts by cryoEM and this revealed multiple, flexible states of the enzyme in adaptation to its association with other complexes, thus exemplifying the need for in situ studies. As the components of the captured protein communities are known—at both the protein and complex levels—this study constitutes another step forward toward a molecular understanding of subcellular organization.
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spelling doaj-art-15f6ec5f9421498ca23ca26e8be3eb202025-08-20T03:46:37ZengSpringer NatureMolecular Systems Biology1744-42922017-07-0113711410.15252/msb.20167412Capturing protein communities by structural proteomics in a thermophilic eukaryotePanagiotis L Kastritis0Francis J O'Reilly1Thomas Bock2Yuanyue Li3Matt Z Rogon4Katarzyna Buczak5Natalie Romanov6Matthew J Betts7Khanh Huy Bui8Wim J Hagen9Marco L Hennrich10Marie‐Therese Mackmull11Juri Rappsilber12Robert B Russell13Peer Bork14Martin Beck15Anne‐Claude Gavin16European Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitCell Networks, Bioquant & Biochemie Zentrum Heidelberg, Heidelberg UniversityEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitChair of Bioanalytics, Institute of Biotechnology, Technische Universität BerlinCell Networks, Bioquant & Biochemie Zentrum Heidelberg, Heidelberg UniversityEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitEuropean Molecular Biology Laboratory, Structural and Computational Biology UnitAbstract The arrangement of proteins into complexes is a key organizational principle for many cellular functions. Although the topology of many complexes has been systematically analyzed in isolation, their molecular sociology in situ remains elusive. Here, we show that crude cellular extracts of a eukaryotic thermophile, Chaetomium thermophilum, retain basic principles of cellular organization. Using a structural proteomics approach, we simultaneously characterized the abundance, interactions, and structure of a third of the C. thermophilum proteome within these extracts. We identified 27 distinct protein communities that include 108 interconnected complexes, which dynamically associate with each other and functionally benefit from being in close proximity in the cell. Furthermore, we investigated the structure of fatty acid synthase within these extracts by cryoEM and this revealed multiple, flexible states of the enzyme in adaptation to its association with other complexes, thus exemplifying the need for in situ studies. As the components of the captured protein communities are known—at both the protein and complex levels—this study constitutes another step forward toward a molecular understanding of subcellular organization.https://doi.org/10.15252/msb.20167412computational modelingcryo‐electron microscopyfatty acid synthaseinteraction proteomicsmetabolon
spellingShingle Panagiotis L Kastritis
Francis J O'Reilly
Thomas Bock
Yuanyue Li
Matt Z Rogon
Katarzyna Buczak
Natalie Romanov
Matthew J Betts
Khanh Huy Bui
Wim J Hagen
Marco L Hennrich
Marie‐Therese Mackmull
Juri Rappsilber
Robert B Russell
Peer Bork
Martin Beck
Anne‐Claude Gavin
Capturing protein communities by structural proteomics in a thermophilic eukaryote
Molecular Systems Biology
computational modeling
cryo‐electron microscopy
fatty acid synthase
interaction proteomics
metabolon
title Capturing protein communities by structural proteomics in a thermophilic eukaryote
title_full Capturing protein communities by structural proteomics in a thermophilic eukaryote
title_fullStr Capturing protein communities by structural proteomics in a thermophilic eukaryote
title_full_unstemmed Capturing protein communities by structural proteomics in a thermophilic eukaryote
title_short Capturing protein communities by structural proteomics in a thermophilic eukaryote
title_sort capturing protein communities by structural proteomics in a thermophilic eukaryote
topic computational modeling
cryo‐electron microscopy
fatty acid synthase
interaction proteomics
metabolon
url https://doi.org/10.15252/msb.20167412
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