Dataset Information

Succination of Tubulin and VDAC2 in the Mouse Brainstem

ABSTRACT: Elevated fumarate concentrations resulting from inhibition of the Krebs cycle lead to increases in protein succination, an irreversible post-translational modification that occurs when fumarate reacts with cysteine residues to generate S-(2-succino)cysteine (2SC). Metabolic events that reduce NADH reoxidation can block Krebs cycle activity, therefore we hypothesized that oxidative phosphorylation deficiencies such as those observed in mitochondrial diseases would also lead to increased protein succination. Using the Ndufs4 knockout (Ndufs4 KO) mouse, a model of Leigh syndrome, we demonstrate for the first time that protein succination is increased in the brainstem (BS), particularly the vestibular nucleus (VN). Notably, the brainstem is the most affected region exhibiting neurodegeneration and astrocyte and microglial proliferation, and these mice typically die of respiratory failure attributed to VN pathology. In contrast, no increases in protein succination were observed in the skeletal muscle, corresponding with the lack of muscle pathology observed in this model. 2D SDS-PAGE followed by immunoblotting for succinated proteins and MS/MS analysis of BS samples allowed us to identify the voltage-dependent anion channels (VDAC) 1 and 2 as specific targets of succination. Using targeted mass spectrometry, Cys77 was identified as a site of endogenous succination in VDAC2. Given the important role of VDAC isoforms in the exchange of ADP/ATP between the cytosol and the mitochondria, and the already decreased capacity for ATP synthesis in the Ndufs4 KO mice, we propose that the increased protein succination observed in the BS of these animals would further decrease the already compromised mitochondrial brain function. These data suggest that fumarate is a novel biochemical link that may contribute to the progression of the neuropathology in this mitochondrial disease model.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain, Neuron

DISEASE(S): Leigh Disease

SUBMITTER: Allison Manuel

LAB HEAD: Norma Frizzell Catledge

PROVIDER: PXD002991 | Pride | 2015-10-16

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Fig2D_2E.msf-pride.pride.mgf.gz	Mgf
	Fig2D_2E.msf-pride.pride.mztab.gz	Mztab
	Fig2D_2E.msf-pride.xml.gz	Xml
	Fig2D_2E.raw	Raw
	Fig5A.msf-pride.pride.mgf.gz	Mgf

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Publications

Succination is Increased on Select Proteins in the Brainstem of the NADH dehydrogenase (ubiquinone) Fe-S protein 4 (Ndufs4) Knockout Mouse, a Model of Leigh Syndrome.

Piroli Gerardo G GG Manuel Allison M AM Clapper Anna C AC Walla Michael D MD Baatz John E JE Palmiter Richard D RD Quintana Albert A Frizzell Norma N

Molecular & cellular proteomics : MCP 20151008 2

Elevated fumarate concentrations as a result of Krebs cycle inhibition lead to increases in protein succination, an irreversible post-translational modification that occurs when fumarate reacts with cysteine residues to generate S-(2-succino)cysteine (2SC). Metabolic events that reduce NADH re-oxidation can block Krebs cycle activity; therefore we hypothesized that oxidative phosphorylation deficiencies, such as those observed in some mitochondrial diseases, would also lead to increased protein ...[more]

PMID: 26450614

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