Manganese ions enhance mitochondrial H2O2 emission from Krebs cycle oxidoreductases by inducing permeability transition

Abstract

Manganese-induced toxicity has been linked to mitochondrial dysfunction and an increased generation of reactive oxygen species (ROS). We could recently show in mechanistic studies that Mn²⁺ ions induce hydrogen peroxide (H₂O₂) production from the ubiquinone binding site of mitochondrial complex II (II_Q) and generally enhance H₂O₂ formation by accelerating the rate of superoxide dismutation. The present study with intact mitochondria reveals that manganese additionally enhances H₂O₂ emission by inducing mitochondrial permeability transition (mPT). In mitochondria fed by NADH-generating substrates, the combination of Mn²⁺ and different respiratory chain inhibitors led to a dynamically increasing H₂O₂emission which was sensitive to the mPT inhibitor cyclosporine A (CsA) as well as Ru-360, an inhibitor of the mitochondrial calcium uniporter (MCU). Under these conditions, flavin-containing enzymes of the mitochondrial matrix, e.g. the mitochondrial 2-oxoglutaratedehydrogenase (OGDH), were major sources of ROS. With succinate as substrate, Mn²⁺ stimulated ROS production mainly at complex II, whereby the applied succinate concentration had a marked effect on the tendency for mPT. Also Ca²⁺ increased the rate of H₂O₂ emission by mPT, while no direct effect on ROS-production of complex II was observed. The present study reveals a complex scenario through which manganese affects mitochondrial H₂O₂ emission: stimulating its production from distinct sites (e.g. site II_Q), accelerating superoxide dismutation and enhancing the emission via mPT which also leads to the loss of soluble components of the mitochondrial antioxidant systems and favors the ROS production from flavin-containing oxidoreductases of the Krebs cycle.

Keywords: 2-oxoglutarate dehydrogenase (OGDH); Complex II (succinate:ubiquinone oxidoreductase); Manganese; Mitochondria; Mitochondrial permeability transition (mPT); Reactive oxygen species (ROS).