The Gpx1 gene encodes mitochondrial glutathione peroxidase in the mouse liver

Mitochondria have GPX and PHGPX activity. It has been an unsettled issue whether mitochondrial GPX is encoded by Gpx1. Unlike the Gpx4 gene which encodes PHGPX with alternative transcription and translation start sites determining the subcellular localization of PHGPX, the Gpx1 gene appears to have a single translation start site. Additionally, mitochondrial GPX has been shown to have different chromatographic and kinetic properties from the cytosolic GPX1. We studied mouse liver mitochondrial GPX activity in homozygous Gpx1-knockout mice. Mitochondria were enriched at the density of 1.10 g/ml in the Percoll gradients as shown by electron microscopy. The H2O2-reducing GPX activity in the highly enriched mitochondrial fraction of wild-type mouse liver is 2700 mU/mg which is about one-half of specific activity found in cytosol. There is less than 0.5% GPX activity in the cytosol and no GPX activity in the mitochondria of Gpx1-knockout mouse liver compared to the cytosol of wild-type mouse liver using H2O2 or cumene hydroperoxide as the substrate. The fact that the knockout mice express normal levels of plasma GPX as well as testis and liver PHGPX activity indicates that animals are not selenium-deficient. Based on these observations, we concluded that mitochondrial GPX is the product of the Gpx1 gene.