Phosphoproteomics of the developing heart identifies PERM1 - An outer mitochondrial membrane protein. (May 2021)

• Record Type: Journal Article
• Authors: Aravamudhan, Sriram; Türk, Clara; Bock, Theresa; Keufgens, Lena; Nolte, Hendrik; Lang, Franziska; Krishnan, Ramesh Kumar; König, Tim; Hammerschmidt, Philipp; Schindler, Natalie; Brodesser, Susanne; Rozsivalova, Dieu Hien; Rugarli, Elena; Trifunovic, Aleksandra; Brüning, Jens; Langer, Thomas; Braun, Thomas; Krüger, Marcus
• Is Part Of: Journal of molecular and cellular cardiology. Volume 154(2021)
• Page Start: 41
• Page End: 59
• Publication Date: 2021-05
• Subjects: Heart development -- Phosphoproteomics -- SILAC -- Mitochondria -- PERM1 -- Lipid metabolism; 4EBP1 Eukaryotic initiation factor 4E binding protein 1 -- ABC Ammonium bicarbonate -- ACN Acetonitrile -- AMD1 S-adenosylmethionine decarboxylase proenzyme 1 -- BAC Bacterial artificial chromosomes -- BN-PAGE Blue native polyacrylamide gel electrophorese -- CaMKII Ca2+/calmodulin-dependent protein kinase -- CK2 Casein kinase 2 -- CLPTM1 Cleft lip and palate transmembrane protein 1 homolog -- DAG Diacylglycerol -- DGC Dystrophin glycoprotein complex -- DTT Dithiothreitol -- ECM Extracellular matrix -- ER Endoplasmic reticulum -- ETS Electron transfer system -- FASP Filter-aided sample preparation -- FCCP Trifluoromethoxyphenylhydrazone -- fld fatty liver dystrophic -- FSTL1 Follistatin-related protein 1 -- H&E Haematoxylin and eosin -- IAA Iodacetamide -- IKK I kappa B kinase -- IT Injection time -- LPIN1 Lipin-1 -- MAM Mitochondria-associated membrane -- MCM Minichromosome maintenance -- MFN2 Mitofusin 2 -- MKL2 Myocardin-related transcription factor B -- NCE Nominal collisional energy -- NMM Non-mitochondrial membrane -- NO Nitric oxide -- PA Phosphatidic acid -- PAP Phosphatidic acid phosphatase -- PE Phosphatidylethanolamine -- PECAM-1 Platelet-endothelial cell adhesion molecule-1 -- PERM1 Peroxisome proliferator–activated receptor γ coactivator 1- and Estrogen-related receptor–induced regulator in muscle 1 -- PF4 Platelet factor 4 -- PI Phosphatidylinositol -- PTM Post-translational modification -- ROX Residual oxygen consumption -- S6K1 Protein S6 kinase -- SCX Strong cation exchange chromatography -- SDH Succinate dehydrogenase -- SILAC Stable isotope labeling with heavy amino acids in cell culture -- SLC Solute carrier family -- TBB 4, 5, 6, 7-tetrabromobenzotriazole -- TFA Trifluoroacetic acid -- TiO2 Titanium dioxide; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2021.01.010
• Languages: English
• ISSNs: 0022-2828

Abstract

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. Graphical abstract: Unlabelled Image Highlights: 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