Mapping hypoxia-induced bioenergetic rearrangement and metabolic signalling by 18O-assisted 31P NMR and 1 NMR spectroscopy

Darko Pucar, Petras P. Dzeja, Peter Bast, Richard J. Gumina, Carmen Drahl, Lynette Lim, Nenad Juranic, Slobodan Macura, Andre Terzic

Research output: Contribution to journalReview article

33 Scopus citations

Abstract

Brief hypoxia or ischemia perturbs energy metabolism inducing paradoxically a stress-tolerant state, yet metabolic signals that trigger cytoprotection remain poorly understood. To evaluate bioenergetic rearrangements, control and hypoxic hearts were analyzed with 18O-assisted 31P NMR and 1H NMR spectroscopy. The 18O-induced isotope shift in the 31P NMR spectrum of CrP, βADP and βATP was used to quantify phosphotransfer fluxes through creatine kinase and adenylate kinase. This analysis was supplemented with determination of energetically relevant metabolites in the phosphomonoester (PME) region of 31P NMR spectra, and in both aromatic and aliphatic regions of 1H NMR spectra. In control conditions, creatine kinase was the major phosphotransfer pathway processing high-energy phosphoryls between sites of ATP consumption and ATP production. In hypoxia, creatine kinase flux was dramatically reduced with a compensatory increase in adenylate kinase flux, which supported heart energetics by regenerating and transferring β- and γ-phosphoryls of ATP. Activation of adenylate kinase led to a build-up of AMP, IMP and adenosine, molecules involved in cardioprotective signaling. 31P and 1H NMR spectral analysis further revealed NADH and H+ scavenging by α-glycerophosphate dehydrogenase (αGPDH) and lactate dehydrogenase contributing to maintained glycolysis under hypoxia. Hypoxia-induced accumulation of α-glycerophosphate and nucleoside 5′-monophosphates, through αGPDH and adenylate kinase reactions, respectively, was mapped within the increased PME signal in the 31P NMR spectrum. Thus, 18O-assisted 31P NMR combined with 1H NMR provide a powerful approach in capturing rearrangements in cardiac bioenergetics, and associated metabolic signaling that underlie the cardiac adaptive response to stress.

Original languageEnglish (US)
Pages (from-to)281-289
Number of pages9
JournalMolecular and Cellular Biochemistry
Volume256-257
Issue number1-2
DOIs
StatePublished - 2004

Keywords

  • Adenylate kinase
  • Creatine kinase
  • Energetics
  • Heart
  • Ischemia
  • Metabolism

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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    Pucar, D., Dzeja, P. P., Bast, P., Gumina, R. J., Drahl, C., Lim, L., Juranic, N., Macura, S., & Terzic, A. (2004). Mapping hypoxia-induced bioenergetic rearrangement and metabolic signalling by 18O-assisted 31P NMR and 1 NMR spectroscopy. Molecular and Cellular Biochemistry, 256-257(1-2), 281-289. https://doi.org/10.1023/b:mcbi.0000009875.30308.7a