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J Proteome Res. 2015 Jan 2;14(1):224-35. doi: 10.1021/pr500894v. Epub 2014 Nov 12.

Metabolome and proteome profiling of complex I deficiency induced by rotenone.

Author information

1
Max Planck Institute for Molecular Genetics , Ihnestra├če 63-73, 14195 Berlin, Germany.

Abstract

Complex I (CI; NADH dehydrogenase) deficiency causes mitochondrial diseases, including Leigh syndrome. A variety of clinical symptoms of CI deficiency are known, including neurodegeneration. Here, we report an integrative study combining liquid chromatography-mass spectrometry (LC-MS)-based metabolome and proteome profiling in CI deficient HeLa cells. We report a rapid LC-MS-based method for the relative quantification of targeted metabolome profiling with an additional layer of confidence by applying multiple reaction monitoring (MRM) ion ratios for further identity confirmation and robustness. The proteome was analyzed by label-free quantification (LFQ). More than 6000 protein groups were identified. Pathway and network analyses revealed that the respiratory chain was highly deregulated, with metabolites such as FMN, FAD, NAD(+), and ADP, direct players of the OXPHOS system, and metabolites of the TCA cycle decreased up to 100-fold. Synthesis of functional iron-sulfur clusters, which are of central importance for the electron transfer chain, and degradation products like bilirubin were also significantly reduced. Glutathione metabolism on the pathway level, as well as individual metabolite components such as NADPH, glutathione (GSH), and oxidized glutathione (GSSG), was downregulated. Overall, metabolome and proteome profiles in CI deficient cells correlated well, supporting our integrated approach.

KEYWORDS:

Complex I deficiency (CI); iron sulfur cluster; mass spectrometry; metabolome profiling; multiple reaction monitoring (MRM); network analysis; pathway analysis; proteome profiling; rotenone

PMID:
25361611
DOI:
10.1021/pr500894v
[Indexed for MEDLINE]

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