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The ‘mitoflash’ probe cpYFP does not respond to superoxide

Author

Listed:
  • Markus Schwarzländer

    (Institute of Crop Science and Resource Conservation (INRES), University of Bonn, Friedrich-Ebert-Allee 144, 53113 Bonn, Germany)

  • Stephan Wagner

    (Institute of Crop Science and Resource Conservation (INRES), University of Bonn, Friedrich-Ebert-Allee 144, 53113 Bonn, Germany)

  • Yulia G. Ermakova

    (Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences)

  • Vsevolod V. Belousov

    (Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences)

  • Rafael Radi

    (and Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, Avda. General Flores 2125, 11800 Montevideo, Uruguay)

  • Joseph S. Beckman

    (Linus Pauling Institute, Environmental Health Sciences Center, Oregon State University)

  • Garry R. Buettner

    (The University of Iowa, and ESR Facility, College of Medicine, Med Labs B180K, Iowa City, Iowa 52242-1181, USA)

  • Nicolas Demaurex

    (University of Geneva, 1, rue Michel-Servet, Geneva 4 CH-1211, Switzerland)

  • Michael R. Duchen

    (University College London, Gower Street, London WC1E 6BT, UK)

  • Henry J. Forman

    (Life and Environmental Sciences Unit, University of California, Merced, 5200 North Lake Road, Merced, California 95344, USA
    Andrus Gerontology Center of the Davis School of Gerontology, University of Southern California, 3715 McClintock Avenue, Los Angeles, California 90089-0191, USA)

  • Mark D. Fricker

    (University of Oxford, South Parks Road, Oxford OX1 3RB, UK)

  • David Gems

    (Institute of Healthy Ageing, Evolution and Environment, University College London, London WC1E 6BT, UK)

  • Andrew P. Halestrap

    (School of Biochemistry and Bristol CardioVascular, University of Bristol, Medical Sciences Building, University Walk, Bristol BS8 1TD, UK)

  • Barry Halliwell

    (National University of Singapore)

  • Ursula Jakob

    (Molecular, University of Michigan)

  • Iain G. Johnston

    (South Kensington Campus, Imperial College London, London SW7 2AZ, UK)

  • Nick S. Jones

    (South Kensington Campus, Imperial College London, London SW7 2AZ, UK)

  • David C. Logan

    (Université d'Angers & INRA & Agrocampus Ouest, UMR 1345 Institut de Recherche en Horticulture et Semences)

  • Bruce Morgan

    (German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany)

  • Florian L. Müller

    (University of Texas MD Anderson Cancer Center)

  • David G. Nicholls

    (Buck Institute for Research on Aging, 8001 Redwood Boulevard)

  • S. James Remington

    (Institute of Molecular Biology, University of Oregon)

  • Paul T. Schumacker

    (Northwestern University, Feinberg School of Medicine, Chicago, Illinois, 60611, USA)

  • Christine C. Winterbourn

    (Centre for Free Radical Research, University of Otago, ChristchurchPO Box 4345, Christchurch, New Zealand)

  • Lee J. Sweetlove

    (University of Oxford, South Parks Road, Oxford OX1 3RB, UK)

  • Andreas J. Meyer

    (Institute of Crop Science and Resource Conservation (INRES), University of Bonn, Friedrich-Ebert-Allee 144, 53113 Bonn, Germany)

  • Tobias P. Dick

    (German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany)

  • Michael P. Murphy

    (MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK)

Abstract

Arising from E.-Z. Shen et al. Nature 508, 128–132 (2014); doi:10.1038/nature1301210.1038/nature13012 Ageing and lifespan of organisms are determined by complicated interactions between their genetics and the environment, but the cellular mechanisms remain controversial; several studies suggest that cellular energy metabolism and free radical dynamics affect lifespan, implicating mitochondrial function. Recently, Shen et al.1 provided apparent mechanistic insight by reporting that mitochondrial oscillations of ‘free radical production’, called ‘mitoflashes’, in the pharynx of three-day old Caenorhabditis elegans correlated inversely with lifespan. The interpretation of mitoflashes as ‘bursts of superoxide radicals’ assumes that circularly permuted yellow fluorescent protein (cpYFP) is a reliable indicator of mitochondrial superoxide2, but this interpretation has been criticized because experiments and theoretical considerations both show that changes in cpYFP fluorescence are due to alterations in pH, not superoxide3,4,5,6,7. Here we show that purified cpYFP is completely unresponsive to superoxide, and that mitoflashes do not reflect superoxide generation or provide a link between mitochondrial free radical dynamics and lifespan. There is a Reply to this Brief Communication Arising by Cheng, H. et al. Nature 514, http://dx.doi.org/10.1038/nature13859 (2014).

Suggested Citation

  • Markus Schwarzländer & Stephan Wagner & Yulia G. Ermakova & Vsevolod V. Belousov & Rafael Radi & Joseph S. Beckman & Garry R. Buettner & Nicolas Demaurex & Michael R. Duchen & Henry J. Forman & Mark D, 2014. "The ‘mitoflash’ probe cpYFP does not respond to superoxide," Nature, Nature, vol. 514(7523), pages 12-14, October.
    • Handle: RePEc:nat:nature:v:514:y:2014:i:7523:d:10.1038_nature13858
    DOI: 10.1038/nature13858
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