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The 'mitoflash' probe cpYFP does not respond to superoxide
Journal article   Peer reviewed

The 'mitoflash' probe cpYFP does not respond to superoxide

Markus Schwarzlaender, Stephan Wagner, Yulia G Ermakova, Vsevolod V Belousov, Rafael Radi, Joseph S Beckman, Garry R Buettner, Nicolas Demaurex, Michael R Duchen, Henry J Forman, …
Nature, Vol.514(7523), pp.E12-E14
2014
DOI: 10.1038/nature13858
PMCID: PMC4346172
PMID: 25341790

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Abstract

Ageing and lifespan of organisms are determined by complicated interactions between their genetics and the environment, but the cellular mechanisms remain controversial. There have been a number of studies suggesting that cellular energy metabolism and free radical dynamics affect lifespan, implicating mitochondrial function. Recently, Shen et al. provided apparent mechanistic insight by reporting that mitochondrial oscillations of ‘free radical production’, called ‘mitoflashes’, in the pharynx of 3-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 superoxide. This interpretation has been criticised because experiments and theoretical considerations both show that changes in cpYFP fluorescence are due to alterations in pH, not superoxide-. We now provide direct evidence that purified cpYFP is completely unresponsive to superoxide. Therefore ‘mitoflashes’ do not reflect superoxide generation and are not evidence for a link between mitochondrial free radical dynamics and lifespan.
Life Sciences

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