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 |
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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. |
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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 – sequence: 5 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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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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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 |
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