PXD060675

PXD060675 is an original dataset announced via ProteomeXchange.

Title	Protein Succinylome analysis identifies citrate synthase as a central regulator of osteoclast metabolic activity
Description	The cytokines tumor necrosis factor ligand superfamily member 11 (TNFSF11; also known as RANKL) and macrophage colony-stimulating factor 1 receptor (M-CSF) differentiate macrophages into bone-resorbing osteoclasts via a process characterised by changes in metabolic activity that support energy-consuming processes such as cell fusion or bone resorption. Treatment with RANKL triggers a phenotype of accelerated metabolism with enhanced glycolysis and an initial disruption of the tricarboxylic acid cycle (TCA) cycle through increased expression of the enzyme aconitate decarboxylase (ACOD1). which results in an upregulation of intracellular succinate levels. Succinate then causes post-translational succinylation of lysine residues. Interestingly, ACOD1 as an inducer of protein succinylation and the desuccinylase NAD-dependent protein deacylase sirtuin-5, mitochondrial (SIRT5) are regulated differentially and the initially high expression of ACOD1 decreases towards the end of differentiation, whereas SIRT5 levels increase. To mimic the effect of protein succinylation, diethyl succinate or a SIRT5 inhibitor were added to differentiating osteoclasts, which reduced the formation of large osteoclasts, showing its relevance for successful osteoclastogenesis. To identify proteins succinylated after RANKL treatment, we used an immunoaffinity-based liquid chromatography–tandem mass spectrometry (LC-MS/MS) approach. Most lysine succinylated proteins were metabolic enzymes localised in the mitochondria. Citrate synthase, the enzyme catalysing the first reaction of the TCA cycle, showed a notable difference in succinylation levels before and after RANKL stimulation, with succinylation detected exclusively in stimulated cells. Immunoprecipitation assays confirmed citrate synthase succinylation. Using whole cell extracts, we observed that RANKL treatment decreased CS activity in a concentration-dependent manner. This suggests that CS could be a critical factor in the context of energy production during osteoclastogenesis and that protein succinylation helps to modulate the differentiation program of osteoclasts.
HostingRepository	PRIDE
AnnounceDate	2025-07-21
AnnouncementXML	Submission_2025-07-20_16:09:25.362.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Marcin Luzarowski
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-11 00:54:11	ID requested
⏵ 1	2025-07-20 16:09:26	announced

10.1111/febs.70090;

Yu D, Gao Y, Luzarowski M, Seebach E, Heitkamp T, B, ö, rsch M, Ruppert T, Kubatzky KF, Protein succinylome analysis identifies citrate synthase as a central regulator of osteoclast metabolic activity. FEBS J, 292(14):3736-3754(2025) [pubmed]

submitter keyword: succinylation, post translational modification,steoclast, citrate synthase, RANKL

Marcin Luzarowski
contact affiliation	Core Facility for Mass Spectrometry and Proteomics, Center for Molecular Biology at Heidelberg University (ZMBH), Im Neuenheimer Feld 345, 69120, Heidelberg, Germany
contact email	m.luzarowski@zmbh.uni-heidelberg.de
lab head
Marcin Luzarowski
contact affiliation	University of Heidelberg
contact email	m.luzarowski@zmbh.uni-heidelberg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/07/PXD060675

PRIDE project URI

• PRIDE
  1. PXD060675
     1. Label: PRIDE project
     2. Name: Protein Succinylome analysis identifies citrate synthase as a central regulator of osteoclast metabolic activity