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- Title
Mitochondrial Metabolomics of Sym1-Depleted Yeast Cells Revealed Them to Be Lysine Auxotroph.
- Authors
Lagies, Simon; Pan, Daqiang; Mohl, Daniel A.; Plattner, Dietmar A.; Gentle, Ian E.; Kammerer, Bernd
- Abstract
Metabolomics has expanded from cellular to subcellular level to elucidate subcellular compartmentalization. By applying isolated mitochondria to metabolome analysis, the hallmark of mitochondrial metabolites has been unraveled, showing compartment-specific distribution and regulation of metabolites. This method was employed in this work to study a mitochondrial inner membrane protein Sym1, whose human ortholog MPV17 is related to mitochondria DNA depletion syndrome. Gas chromatography–mass spectrometry-based metabolic profiling was combined with targeted liquid chromatography–mass spectrometry analysis to cover more metabolites. Furthermore, we applied a workflow employing ultra-high performance liquid chromatography–quadrupole time of flight mass spectrometry with a powerful chemometrics platform, focusing on only significantly changed metabolites. This workflow highly reduced the complexity of acquired data without losing metabolites of interest. Consequently, forty-one novel metabolites were identified in addition to the combined method, of which two metabolites, 4-guanidinobutanal and 4-guanidinobutanoate, were identified for the first time in Saccharomyces cerevisiae. With compartment-specific metabolomics, we identified sym1Δ cells as lysine auxotroph. The highly reduced carbamoyl-aspartate and orotic acid indicate a potential role of the mitochondrial inner membrane protein Sym1 in pyrimidine metabolism.
- Subjects
METABOLOMICS; TIME-of-flight mass spectrometry; LIQUID chromatography-mass spectrometry; OROTIC acid
- Publication
Cells (2073-4409), 2023, Vol 12, Issue 5, p692
- ISSN
2073-4409
- Publication type
Article
- DOI
10.3390/cells12050692