Leishmania tarentolae: A parallel isolation of cytochrome bc(1) and cytochrome c oxidase

A rapid and simple method which allowed for a parallel isolation of cytochr ome c reductase (cytochrome bell and cytochrome c oxidase from kinetoplast- mitochondria of Leishmania tarentolae was developed. The method involved th e lysis of kinetoplasts with dodecyl maltoside in the presence of 260 mM Na Cl, followed by purification of bc(1) complexes on DEAE-sepharose CL-6B. Th e oxidase which was found in the flow-through fractions of the first chroma tographic step was diluted and then repurified on a similar DEAE-sepharose column. The investigated properties of the isolated cytochrome c oxidase an d reductase, such as their absolute and difference spectrum absorption maxi ma, heme content, specific activity, and subunit composition, confirm the u sefulness of this method for obtaining highly active preparations of the en zymes. (C) 2000 Academic Press.