Using exomarkers to assess mitochondrial reactive species in vivo

• Published in: Biochimica et biophysica acta Vol. 1840; no. 2; pp. 923 - 930
• Main Authors: Logan, Angela, Cochemé, Helena M., Li Pun, Pamela Boon, Apostolova, Nadezda, Smith, Robin A.J., Larsen, Lesley, Larsen, David S., James, Andrew M., Fearnley, Ian M., Rogatti, Sebastian, Prime, Tracy A., Finichiu, Peter G., Dare, Anna, Chouchani, Edward T., Pell, Victoria R., Methner, Carmen, Quin, Caroline, McQuaker, Stephen J., Krieg, Thomas, Hartley, Richard C., Murphy, Michael P.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2014
• Subjects: Animals; biomarkers; Biomarkers - analysis; biophysics; Diptera; Exomarker; hydrogen peroxide; Mass spectrometry; membrane proteins; Mice; MitoB; Mitochondria; Mitochondria - metabolism; Models, Biological; Molecular Probes; Oxidative damage; Oxidative Stress; Reactive oxygen species; Reactive Oxygen Species - analysis; Reactive oxygen species; MitoB; Oxidative damage; EPR; SOD; Exomarker; GFP; Mitochondria; 4-HNE; TPP; MitoP; ROS; TPMP; Mass spectrometry; triphenylphosphonium cation; 3-(dihydroxyboronyl)benzyltriphenylphosphonium bromide; methyltriphenylphosphonium; green fluorescent protein; 4-hydroxynonenal; (3-hydroxybenzyl)triphenylphosphonium bromide; electron paramagnetic resonance; superoxide dismutase
• DOI: 10.1016/j.bbagen.2013.05.026
• ISSN: 0304-4165; 0006-3002

The ability to measure the concentrations of small damaging and signalling molecules such as reactive oxygen species (ROS) in vivo is essential to understanding their biological roles. While a range of methods can be applied to in vitro systems, measuring the levels and relative changes in reactive species in vivo is challenging. One approach towards achieving this goal is the use of exomarkers. In this, exogenous probe compounds are administered to the intact organism and are then transformed by the reactive molecules in vivo to produce a diagnostic exomarker. The exomarker and the precursor probe can be analysed ex vivo to infer the identity and amounts of the reactive species present in vivo. This is akin to the measurement of biomarkers produced by the interaction of reactive species with endogenous biomolecules. Our laboratories have developed mitochondria-targeted probes that generate exomarkers that can be analysed ex vivo by mass spectrometry to assess levels of reactive species within mitochondria in vivo. We have used one of these compounds, MitoB, to infer the levels of mitochondrial hydrogen peroxide within flies and mice. Here we describe the development of MitoB and expand on this example to discuss how better probes and exomarkers can be developed. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn. •Exomarkers can be used to assess reactive species in vivo.•MitoB is a mitochondria-targeted ROS probe.•Tandem mass spectrometry can be used to assess ROS in vivo.