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2018 ; 14
(5
): 59
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Metabolic profiling of isolated mitochondria and cytoplasm reveals
compartment-specific metabolic responses
#MMPMID29628813
Pan D
; Lindau C
; Lagies S
; Wiedemann N
; Kammerer B
Metabolomics
2018[]; 14
(5
): 59
PMID29628813
show ga
INTRODUCTION: Subcellular compartmentalization enables eukaryotic cells to carry
out different reactions at the same time, resulting in different metabolite pools
in the subcellular compartments. Thus, mutations affecting the mitochondrial
energy metabolism could cause different metabolic alterations in mitochondria
compared to the cytoplasm. Given that the metabolite pool in the cytosol is
larger than that of other subcellular compartments, metabolic profiling of total
cells could miss these compartment-specific metabolic alterations. OBJECTIVES: To
reveal compartment-specific metabolic differences, mitochondria and the
cytoplasmic fraction of baker's yeast Saccharomyces cerevisiae were isolated and
subjected to metabolic profiling. METHODS: Mitochondria were isolated through
differential centrifugation and were analyzed together with the remaining
cytoplasm by gas chromatography-mass spectrometry (GC-MS) based metabolic
profiling. RESULTS: Seventy-two metabolites were identified, of which eight were
found exclusively in mitochondria and sixteen exclusively in the cytoplasm. Based
on the metabolic signature of mitochondria and of the cytoplasm, mutants of the
succinate dehydrogenase (respiratory chain complex II) and of the
F(O)F(1)-ATP-synthase (complex V) can be discriminated in both compartments by
principal component analysis from wild-type and each other. These mitochondrial
oxidative phosphorylation machinery mutants altered not only citric acid cycle
related metabolites but also amino acids, fatty acids, purine and pyrimidine
intermediates and others. CONCLUSION: By applying metabolomics to isolated
mitochondria and the corresponding cytoplasm, compartment-specific metabolic
signatures can be identified. This subcellular metabolomics analysis is a
powerful tool to study the molecular mechanism of compartment-specific metabolic
homeostasis in response to mutations affecting the mitochondrial metabolism.
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