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 inJournal of molecular and cellular cardiology Vol. 154; pp. 41 - 59
Main 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
Format Journal Article
LanguageEnglish
Published England Elsevier Ltd 01.05.2021
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Online AccessGet full text
ISSN0022-2828
1095-8584
1095-8584
DOI10.1016/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. [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
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
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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
Language English
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Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.
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Snippet Heart development relies on PTMs that control cardiomyocyte proliferation, differentiation and cardiac morphogenesis. We generated a map of phosphorylation...
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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
URI https://www.clinicalkey.com/#!/content/1-s2.0-S0022282821000250
https://dx.doi.org/10.1016/j.yjmcc.2021.01.010
https://www.ncbi.nlm.nih.gov/pubmed/33549681
https://www.proquest.com/docview/2487429097
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