The interactome of intact mitochondria by cross-linking mass spectrometry provides evidence for coexisting respiratory supercomplexes

Mitochondria exert an immense amount of cytophysiological functions, but the structural basis of most of these processes is still poorly understood. Here we use cross-linking mass spectrometry to probe the organization of proteins in native mouse heart mitochondria. Our approach provides the largest...

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Published in:Molecular & cellular proteomics Vol. 17; no. 2; pp. 216 - 232
Main Authors: Liu, Fan, Lössl, Philip, Rabbitts, Beverley M., Balaban, Robert S., Heck, Albert J.R.
Format: Journal Article
Language:English
Published: United States Elsevier Inc 01-02-2018
American Society for Biochemistry and Molecular Biology
The American Society for Biochemistry and Molecular Biology
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Abstract Mitochondria exert an immense amount of cytophysiological functions, but the structural basis of most of these processes is still poorly understood. Here we use cross-linking mass spectrometry to probe the organization of proteins in native mouse heart mitochondria. Our approach provides the largest survey of mitochondrial protein interactions reported so far. In total, we identify 3,322 unique residue-to-residue contacts involving half of the mitochondrial proteome detected by bottom-up proteomics. The obtained mitochondrial protein interactome gives insights in the architecture and submitochondrial localization of defined protein assemblies, and reveals the mitochondrial localization of four proteins not yet included in the MitoCarta database. As one of the highlights, we show that the oxidative phosphorylation complexes I-V exist in close spatial proximity, providing direct evidence for supercomplex assembly in intact mitochondria. The specificity of these contacts is demonstrated by comparative analysis of mitochondria after high salt treatment, which disrupts the native supercomplexes and substantially changes the mitochondrial interactome.
AbstractList Mitochondria exert an immense amount of cytophysiological functions, but the structural basis of most of these processes is still poorly understood. Here we use cross-linking mass spectrometry to probe the organization of proteins in native mouse heart mitochondria. Our approach provides the largest survey of mitochondrial protein interactions reported so far. In total, we identify 3,322 unique residue-to-residue contacts involving half of the mitochondrial proteome detected by bottom-up proteomics. The obtained mitochondrial protein interactome gives insights in the architecture and submitochondrial localization of defined protein assemblies, and reveals the mitochondrial localization of four proteins not yet included in the MitoCarta database. As one of the highlights, we show that the oxidative phosphorylation complexes I-V exist in close spatial proximity, providing direct evidence for supercomplex assembly in intact mitochondria. The specificity of these contacts is demonstrated by comparative analysis of mitochondria after high salt treatment, which disrupts the native supercomplexes and substantially changes the mitochondrial interactome.
Author Balaban, Robert S.
Heck, Albert J.R.
Lössl, Philip
Liu, Fan
Rabbitts, Beverley M.
Author_xml – sequence: 1
  givenname: Fan
  surname: Liu
  fullname: Liu, Fan
  organization: From the Biomolecular Mass Spectrometry and Proteomics. Bijvoet Centre for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH, Utrecht, The Netherlands
– sequence: 2
  givenname: Philip
  surname: Lössl
  fullname: Lössl, Philip
  organization: From the Biomolecular Mass Spectrometry and Proteomics. Bijvoet Centre for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH, Utrecht, The Netherlands
– sequence: 3
  givenname: Beverley M.
  surname: Rabbitts
  fullname: Rabbitts, Beverley M.
  organization: Laboratory of Cardiac Energetics, Systems Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD
– sequence: 4
  givenname: Robert S.
  surname: Balaban
  fullname: Balaban, Robert S.
  email: balabanr@nhlbi.nih.gov
  organization: Laboratory of Cardiac Energetics, Systems Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD
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  givenname: Albert J.R.
  surname: Heck
  fullname: Heck, Albert J.R.
  email: a.j.r.heck@uu.nl
  organization: From the Biomolecular Mass Spectrometry and Proteomics. Bijvoet Centre for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH, Utrecht, The Netherlands
BackLink https://www.ncbi.nlm.nih.gov/pubmed/29222160$$D View this record in MEDLINE/PubMed
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ContentType Journal Article
Copyright 2018 © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
2018 by The American Society for Biochemistry and Molecular Biology, Inc.
Copyright American Society for Biochemistry and Molecular Biology Feb 2018
2018 by The American Society for Biochemistry and Molecular Biology, Inc. 2018 The American Society for Biochemistry and Molecular Biology, Inc.
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Notes These authors contributed equally to this work.
Author contributions: F.L., P.L., B.M.R., R.S.B., and A.J.R.H. designed the research; F.L., P.L., and B.M.R. performed the research; F.L. and B.M.R. contributed new reagents/analytic tools; F.L., P.L., R.S.B., and A.J.R.H. analyzed data; and F.L., P.L., B.M.R., R.S.B., and A.J.R.H. wrote the paper.
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Snippet Mitochondria exert an immense amount of cytophysiological functions, but the structural basis of most of these processes is still poorly understood. Here we...
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proquest
crossref
pubmed
elsevier
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StartPage 216
SubjectTerms Animals
Comparative analysis
Crosslinking
Localization
Male
Mass Spectrometry
Mass spectroscopy
Mice, Inbred C57BL
Mitochondria
Mitochondria, Heart - drug effects
Mitochondria, Heart - metabolism
Mitochondrial Proteins - metabolism
Multiprotein Complexes - metabolism
Oxidative phosphorylation
Phosphorylation
Protein interaction
Protein Interaction Maps
Proteins
Proteomics
Salts
Scientific imaging
Sodium Chloride - pharmacology
Spectroscopy
Title The interactome of intact mitochondria by cross-linking mass spectrometry provides evidence for coexisting respiratory supercomplexes
URI https://dx.doi.org/10.1074/mcp.RA117.000470
https://www.ncbi.nlm.nih.gov/pubmed/29222160
https://www.proquest.com/docview/2006902915
https://pubmed.ncbi.nlm.nih.gov/PMC5795388
Volume 17
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