Expression and purification of proteins involved in the mitochondrial electron transport chain

Abstract

NADH: ubiquinone oxidoredutase is one of the most intricate enzymes. It is part of the respiratory chains and is present in several organisms. This enzyme couples the transfer of two electron, from NADH to ubiquinone, and translocate ions across the membrane establishing a transmembrane difference of electrochemical potential. In this work, we present the initial experimental steps to express and purify a subunit from this intricate enzyme, which is directly related with the translocation of ions (i.e. NuoL). The objective was to characterize the subunit reconstituted in membrane-mimicking nanoparticles called nanodiscs using solution Nuclear Magnetic Resonance spectroscopy.