Phosphoproteomics of the developing heart identifies PERM1 - An outer mitochondrial membrane protein
Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that w...
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Published in | Journal of molecular and cellular cardiology Vol. 154; pp. 41 - 59 |
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Main Authors | , , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
Elsevier Ltd
01.05.2021
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Subjects | |
Online Access | Get full text |
ISSN | 0022-2828 1095-8584 1095-8584 |
DOI | 10.1016/j.yjmcc.2021.01.010 |
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Abstract | Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1−/− mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria.
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•Proteomes from mouse hearts during early post-natal development cover >10,000 phosphorylation sites on 5000 proteins.•Skeletal muscle and heart specific PGC-1- and ERR-induced regulator in muscle 1 (PERM1) expression increases from P2 to P20•PERM1 associates with the outer mitochondrial membrane•PERM1 is regulated by phosphorylation on its PEST domain by casein kinase 2•Ablation of Perm1 alters levels of various lipid species, amino acids, and acylcarnitines |
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AbstractList | Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1−/− mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria.
[Display omitted]
•Proteomes from mouse hearts during early post-natal development cover >10,000 phosphorylation sites on 5000 proteins.•Skeletal muscle and heart specific PGC-1- and ERR-induced regulator in muscle 1 (PERM1) expression increases from P2 to P20•PERM1 associates with the outer mitochondrial membrane•PERM1 is regulated by phosphorylation on its PEST domain by casein kinase 2•Ablation of Perm1 alters levels of various lipid species, amino acids, and acylcarnitines Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1 mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria. Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1-/- mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria.Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation sites during the early stages of cardiac postnatal development in mice; we quantified over 10,000 phosphorylation sites and 5000 proteins that were assigned to different pathways. Analysis of mitochondrial proteins led to the identification of PGC-1- and ERR-induced regulator in muscle 1 (PERM1), which is specifically expressed in skeletal muscle and heart tissue and associates with the outer mitochondrial membrane. We demonstrate PERM1 is subject to rapid changes mediated by the UPS through phosphorylation of its PEST motif by casein kinase 2. Ablation of Perm1 in mice results in reduced protein expression of lipin-1 accompanied by accumulation of specific phospholipid species. Isolation of Perm1-deficient mitochondria revealed significant downregulation of mitochondrial transport proteins for amino acids and carnitines, including SLC25A12/13/29/34 and CPT2. Consistently, we observed altered levels of various lipid species, amino acids, and acylcarnitines in Perm1-/- mitochondria. We conclude that the outer mitochondrial membrane protein PERM1 regulates homeostasis of lipid and amino acid metabolites in mitochondria. |
Author | Braun, Thomas Rugarli, Elena Trifunovic, Aleksandra Krüger, Marcus Rozsivalova, Dieu Hien Bock, Theresa Türk, Clara Krishnan, Ramesh Kumar Brodesser, Susanne Brüning, Jens König, Tim Hammerschmidt, Philipp Aravamudhan, Sriram Lang, Franziska Langer, Thomas Schindler, Natalie Nolte, Hendrik Keufgens, Lena |
Author_xml | – sequence: 1 givenname: Sriram surname: Aravamudhan fullname: Aravamudhan, Sriram organization: Cell Signaling Technology, Dellaertweg 9b, 2316 WZ Leiden, Netherlands – sequence: 2 givenname: Clara surname: Türk fullname: Türk, Clara organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 3 givenname: Theresa surname: Bock fullname: Bock, Theresa organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 4 givenname: Lena surname: Keufgens fullname: Keufgens, Lena organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 5 givenname: Hendrik surname: Nolte fullname: Nolte, Hendrik organization: Max Planck Institute for Biology of Ageing, 50931 Cologne, Germany – sequence: 6 givenname: Franziska surname: Lang fullname: Lang, Franziska organization: TRON - Translational Oncology at the University Medical Center of the Johannes Gutenberg University Mainz, 55131 Mainz, Germany – sequence: 7 givenname: Ramesh Kumar surname: Krishnan fullname: Krishnan, Ramesh Kumar organization: Excellence Cluster Cardio-Pulmonary Institute (CPI), Aulweg 130, 35392 Giessen, Germany – sequence: 8 givenname: Tim surname: König fullname: König, Tim organization: Montreal Neurological Institute, McGill University, 3801 University Street, H3A 2B4 Montreal, QC, Canada – sequence: 9 givenname: Philipp surname: Hammerschmidt fullname: Hammerschmidt, Philipp organization: Department of Neuronal Control of Metabolism, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany – sequence: 10 givenname: Natalie surname: Schindler fullname: Schindler, Natalie organization: Institut für Entwicklungsbiologie und Neurobiologie (IDN), Fachbereich Biologie (FB 10), Johannes Gutenberg University (JGU) Mainz, Germany c/o Institute of Molecular Biology gGmbH (IMB), Ackermannweg 4, 55128 Mainz, Germany – sequence: 11 givenname: Susanne surname: Brodesser fullname: Brodesser, Susanne organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 12 givenname: Dieu Hien surname: Rozsivalova fullname: Rozsivalova, Dieu Hien organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 13 givenname: Elena surname: Rugarli fullname: Rugarli, Elena organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 14 givenname: Aleksandra surname: Trifunovic fullname: Trifunovic, Aleksandra organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany – sequence: 15 givenname: Jens surname: Brüning fullname: Brüning, Jens organization: Department of Neuronal Control of Metabolism, Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany – sequence: 16 givenname: Thomas surname: Langer fullname: Langer, Thomas organization: Max Planck Institute for Biology of Ageing, 50931 Cologne, Germany – sequence: 17 givenname: Thomas surname: Braun fullname: Braun, Thomas organization: Max Planck Institute for Heart and Lung Research, Ludwigstr. 43, 61231 Bad Nauheim, Germany – sequence: 18 givenname: Marcus surname: Krüger fullname: Krüger, Marcus email: marcus.krueger@uni-koeln.de organization: CECAD Research Center, Institute for Genetics, University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany |
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Keywords | ETS AMD1 SCX SLC TFA 4EBP1 Mitochondria TBB SDH Lipid metabolism MKL2 DGC Heart development H&E IAA IT ER NCE MAM ROX TiO2 NO PECAM-1 ABC DTT PERM1 PTM FCCP CK2 FSTL1 CLPTM1 MFN2 fld Phosphoproteomics BAC BN-PAGE IKK SILAC DAG ACN ECM NMM S6K1 PA CaMKII PE MCM FASP PI LPIN1 PAP PF4 |
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SubjectTerms | Heart development Lipid metabolism Mitochondria PERM1 Phosphoproteomics SILAC |
Title | Phosphoproteomics of the developing heart identifies PERM1 - An outer mitochondrial membrane protein |
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