title: Decoding the dynamics of cellular metabolism and the action of 3-bromopyruvate and 2-deoxyglucose using pulsed stable isotope-resolved metabolomics.
abstract: BACKGROUND
  Cellular metabolism is highly dynamic and continuously adjusts to the physiological program of the cell. The regulation of metabolism appears at all biological levels: (post-) transcriptional, (post-) translational, and allosteric. This regulatory information is expressed in the metabolome, but in a complex manner. To decode such complex information, new methods are needed in order to facilitate dynamic metabolic characterization at high resolution.
  
  
  RESULTS
  Here, we describe pulsed stable isotope-resolved metabolomics (pSIRM) as a tool for the dynamic metabolic characterization of cellular metabolism. We have adapted gas chromatography-coupled mass spectrometric methods for metabolomic profiling and stable isotope-resolved metabolomics. In addition, we have improved robustness and reproducibility and implemented a strategy for the absolute quantification of metabolites.
  
  
  CONCLUSIONS
  By way of examples, we have applied this methodology to characterize central carbon metabolism of a panel of cancer cell lines and to determine the mode of metabolic inhibition of glycolytic inhibitors in times ranging from minutes to hours. Using pSIRM, we observed that 2-deoxyglucose is a metabolic inhibitor, but does not directly act on the glycolytic cascade.
authors.affiliation: Integrative Metabolomics and Proteomics, Berlin Institute of Medical Systems Biology/Max-Delbrueck Center for Molecular Medicine, Robert Rossle Street 10, Berlin 13125, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Pietzke
authors.affiliation: Integrative Metabolomics and Proteomics, Berlin Institute of Medical Systems Biology/Max-Delbrueck Center for Molecular Medicine, Robert Rossle Street 10, Berlin 13125, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Zasada
authors.affiliation: Integrative Metabolomics and Proteomics, Berlin Institute of Medical Systems Biology/Max-Delbrueck Center for Molecular Medicine, Robert Rossle Street 10, Berlin 13125, Germany.|Present address: Faculté des Sciences, de la Technologie et de la Communication, University of Luxembourg, 162 A, Avenue de la Faïencerie L-1511, Luxembourg, Luxembourg.
authors.forename: Susann
authors.initials: S
authors.lastname: Mudrich
authors.affiliation: Integrative Metabolomics and Proteomics, Berlin Institute of Medical Systems Biology/Max-Delbrueck Center for Molecular Medicine, Robert Rossle Street 10, Berlin 13125, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kempa
keywords: 2-Deoxyglucose; 3-Bromopyruvate; Cancer metabolism; GC-MS; Glycolysis; Metabolomics; Stable isotope labeling