Fulltext

Phosphoproteome analysis of isoflurane-protected heart mitochondria: phosphorylation of adenine nucleotide translocator-1 on Tyr194 regulates mitochondrial function

Feng, Jianhua ; Zhu, Min ; Schaub, Marcus C. ; Gehrig, Peter ; Roschitzki, Bernd ; Lucchinetti, Eliana ; Zaugg, Michael

In: Cardiovascular Research, 2008, vol. 80, no. 1, p. 20-29

Zum persönliche Liste hinzufügen
    Titel
    • Phosphoproteome analysis of isoflurane-protected heart mitochondria: phosphorylation of adenine nucleotide translocator-1 on Tyr194 regulates mitochondrial function
    Autor
    • Feng, Jianhua. Cardiovascular Anesthesia Research Laboratory, Institute of Anesthesiology, E-HOF, University Hospital Zurich, Switzerland, and Center of Integrative Human Physiology, University of Zurich, Rämistrasse 100, CH-8091 Zurich, Switzerland
    • Zhu, Min. Cardiovascular Anesthesia Research Laboratory, Institute of Anesthesiology, E-HOF, University Hospital Zurich, Switzerland, and Center of Integrative Human Physiology, University of Zurich, Rämistrasse 100, CH-8091 Zurich, Switzerland
    • Schaub, Marcus C.. Institute of Pharmacology and Toxicology, University of Zurich, Switzerland
    • Gehrig, Peter. Functional Genomics Center Zurich, Swiss Federal Institute of Technology Zurich and University of Zurich, Switzerland
    • Roschitzki, Bernd. Functional Genomics Center Zurich, Swiss Federal Institute of Technology Zurich and University of Zurich, Switzerland
    • Lucchinetti, Eliana. Cardiovascular Anesthesia Research Laboratory, Institute of Anesthesiology, E-HOF, University Hospital Zurich, Switzerland, and Center of Integrative Human Physiology, University of Zurich, Rämistrasse 100, CH-8091 Zurich, Switzerland
    • Zaugg, Michael. Cardiovascular Anesthesia Research Laboratory, Institute of Anesthesiology, E-HOF, University Hospital Zurich, Switzerland, and Center of Integrative Human Physiology, University of Zurich, Rämistrasse 100, CH-8091 Zurich, Switzerland
    Dokumententyp
    • Postprint
    Sprache
    • Englisch
    Veröffentlicht in
    • Cardiovascular Research, 2008, vol. 80, no. 1, p. 20-29. Oxford University Press
    Andere elektronische Ausgabe
    OAI-PMH-ID
    • oai:doc.rero.ch:302877
    Summary
    Aims Reversible phosphorylation of mitochondrial proteins is essential in the regulation of respiratory function, energy metabolism, and mitochondrion-mediated cell death. We hypothesized that mitochondrial protein phosphorylation plays a critical role in cardioprotection during pre and postconditioning, two of the most efficient anti-ischaemic therapies. Methods and results Using phosphoproteomic approaches, we investigated the profiles of phosphorylated proteins in Wistar rat heart mitochondria protected by pharmacological pre and postconditioning elicited by isoflurane. Sixty-one spots were detected by two-dimensional blue-native gel electrophoresis-coupled Western blotting using a phospho-Ser/Thr/Tyr-specific antibody, and 45 of these spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Eleven protein spots related to oxidative phosphorylation, energy metabolism, chaperone, and carrier functions exhibited significant changes in their phosphorylation state when protected mitochondria were compared with unprotected. Using a phosphopeptide enrichment protocol followed by liquid chromatography-MS/MS, 26 potential phosphorylation sites were identified in 19 proteins. Among these, a novel phosphorylation site was detected in adenine nucleotide translocator-1 (ANT1) at residue Tyr194. Changes in ANT phosphorylation between protected and unprotected mitochondria were confirmed by immunoprecipitation. The biological significance of ANT phosphorylation at Tyr194 was further tested with site-directed mutagenesis in yeast. Substitution of Tyr194 with Phe, mimicking the non-phosphorylated state, resulted in the inhibition of yeast growth on non-fermentable carbon sources, implying a critical role of phosphorylation at this residue in regulating ANT function and cellular respiration. Conclusions Our analysis emphasizes the regulatory functions of the phosphoproteome in heart mitochondria and reveals a novel, potential link between bioenergetics and cardioprotection