Related Experiment Video
Updated: Jun 27, 2026

Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays
Published on: February 10, 2022
Restoration of electron transport without proton pumping in mammalian mitochondria
Ester Perales-Clemente1, Maria Pilar Bayona-Bafaluy, Acisclo Pérez-Martos
1Departamento de Bioquímica. Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain.
Abstract:
We have restored the CoQ oxidative capacity of mouse mtDNA-less cells (rho degrees cells) by transforming them with the alternative oxidase Aox of Emericella nidulans. Cotransforming rho degrees cells with the NADH dehydrogenase of Saccharomyces cerevisiae, Ndi1 and Aox recovered the NADH DH/CoQ reductase and the CoQ oxidase activities. CoQ oxidation by AOX reduces the dependence of rho degrees cells on pyruvate and uridine. Coexpression of AOX and NDI1 further improves the recycling of NAD(+). Therefore, 2 single-protein enzymes restore the electron transport in mammalian mitochondria substituting >80 nuclear DNA-encoded and 11 mtDNA-encoded proteins. Because those enzymes do not pump protons, we were able to split electron transport and proton pumping (ATP synthesis) and inquire which of the metabolic deficiencies associated with the loss of oxidative phosphorylation should be attributed to each of the 2 processes.
Related Concept Videos
Chemiosmosis and ATP Synthesis
Electron Transport Chain Components
Electron Transport Chains
The ETC is comprised of...
Electron Transport Chain: Complex III and IV
The Electron Transport Chain
Inhibitors of the electron transport chain
Rotenone, a widely used pesticide, prevents electron transfer from Fe-S cluster to ubiquinone or Q in...
Chemiosmosis
Electron Transport Chain
The electron transport chain involves a series of protein complexes on the inner mitochondrial membrane that undergo a series of redox reactions. At the end of this chain, the electrons reduce...

