Topology of Superoxide Production from Different Sites in the Mitochondrial Electron Transport Chain*

By J. St-Pierre, Julie A. Buckingham, Stephen J Roebuck, M. Brand. Journal of Biological Chemistry, 2002

1,548 citations recorded by the source corpus.

We measured production of reactive oxygen species by intact mitochondria from rat skeletal muscle, heart, and liver under various experimental conditions. By using different substrates and inhibitors, we determined the sites of production (which complexes in the electron transport chain produced superoxide). By measuring hydrogen peroxide production in the absence and presence of exogenous superoxide dismutase, we established the topology of superoxide production (on which side of the mitochondrial inner membrane superoxide was produced). Mitochondria did not release measurable amounts of superoxide or hydrogen peroxide when respiring on complex I or complex II substrates. Mitochondria from skeletal muscle or heart generated significant amounts of superoxide from complex I when respiring on palmitoyl carnitine. They produced superoxide at considerable rates in the presence of various inhibitors of the electron transport chain. Complex I (and perhaps the fatty acid oxidation electron transfer flavoprotein and its oxidoreductase) released superoxide on the matrix side of the inner membrane, whereas center o of complex III released superoxide on the cytoplasmic side. These results do not support the idea that mitochondria produce considerable amounts of reactive oxygen species under physiological conditions. Our upper estimate of the proportion of electron flow giving rise to hydrogen peroxide with palmitoyl carnitine as substrate (0.15%) is more than an order of magnitude lower than commonly cited values. We observed no difference in the rate of hydrogen peroxide production between rat and pigeon heart mitochondria respiring on complex I substrates. However, when complex I was fully reduced using rotenone, rat mitochondria released significantly more hydrogen peroxide than pigeon mitochondria. This difference was solely due to an elevated concentration of complex I in rat compared with pigeon heart mitochondria.

Subfields: Molecular Biology, Physiology

View Paper: https://www.semanticscholar.org/paper/eaf5d2909929cc1f57f2c345834e8aaa5de83468

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

1. R. Kaufman
2. J. Malhotra
3. J. Speakman
4. M. Juhaszova
5. S. S. Cao
6. S. Dikalov
7. M. Brand
8. D. Zorov

In Physiology

1. J. Speakman
2. M. Brand
3. D. Costantini
4. R. Kaufman
5. W. Buttemer
6. J. Malhotra
7. W. Sivitz
8. M. Yorek

Citations in the corpus

• Exercise-Induced Oxidative Stress: Cellular Mechanisms and Impact on Muscle Force Production (2008, neutral): “Brand and colleagues (371) have recently reassessed”
• Mitochondria in health and disease: perspectives on a new mitochondrial biology. (2004, background): “Recent evidence suggests strongly that free radicals seem to be generated from complex I through oxidation of NADH on the matrix side of the inner mitochondrial membrane (St-Pierre et al., 2002).”
• Molecular Mechanisms of Ischemia-Reperfusion Injury in Brain: Pivotal Role of the Mitochondrial Membrane Potential in Reactive Oxygen Species Generation (2012, background): “This ROS production takes place proximal to CcO, at complexes I and III, which release superoxide on the matrix and intermembrane space sides, respectively [68].”
• Calcium, ATP, and ROS: a mitochondrial love-hate triangle. (2004, background): “Complex I is also a source of ROS, although the mechanism of generation is less clear than for complex III.”
• Endoplasmic Reticulum Stress and Oxidative Stress in Cell Fate Decision and Human Disease (2014, background): “Mitochondrial Ca also activates nitric oxide synthase, whose product disturbs the electron transport chain and enhances ROS generation (16,159).”
• How mitochondria produce reactive oxygen species (2008, background): “…2 •− production, [E], will vary with organism, tissue, state, age or hormonal status, and may underlie many of the changes in maximum ROS production capacity between tissues [23]; for example, complex I content may explain the different maximum capacities of pigeon and rat heart mitochondria [24].”
• The mitochondrial energy transduction system and the aging process. (2007, neutral): “15% with palmitoyl carnitine as substrate (148).”
• Body size, energy metabolism and lifespan (2005, background): “When the membrane potential is low, mitochondria produce very few ROS (St Pierre et al., 2002), suggesting that production from complex 1 is normally minor, or is similarly dependent on membrane potential.”
• Regulation of reactive oxygen species generation in cell signaling (2011, background): “In response to external stresses, p66Shc is phosphorylated on serine 36 residue by the stress-activated kinase PKCbeta and translocated into the mitochondrial IMS (Gertz et al.,”
• The sites and topology of mitochondrial superoxide production (2010, background): “…III produces superoxide at high rates only in the presence of antimycin (Chen et al., 2003; Gyulkhandanyan and Pennefather, 2004; Herrero and Barja, 1997; Kwong and Sohal, 1998; Liu et al., 2002; McLennan and Degli Esposti, 2000; Raha et al., 2000; St-Pierre et al., 2002; Turrens et al., 1985).”