Functional alteration of cytochrome c oxidase by SURF1 mutations in Leigh syndrome

Subacute necrotising encephalomyopathy (Leigh syndrome) due to cytochrome c oxidase (COX) deficiency is often caused by mutations in the SURFl gene, encoding the Surfl protein essential for COX assembly. We have investigated five patients with different SURFl mutations resulting in the absence of Surfl protein. All of them presented with severe and generalised COX defect. Immunoelectrophoretic analysis of cultured fibroblasts revealed 85% decrease of the normal-size COX complexes and significant accumulation of incomplete COX assemblies of 90-120 kDa. Spectrophotometric assay of COX activity showed a 70-90% decrease in lauryl maltoside (LM)-solubilised fibroblasts. In contrast, oxygen consumption analysis in whole cells revealed only a 13-31% decrease of COX activity, which was completely inhibited by detergent in patient cells but not in controls. In patient fibroblasts ADP-stimulated respiration was 50% decreased and cytofluorometry showed a significant decrease of mitochondrial membrane potential Deltapsi(m) in state 4, as well as a 2.4-fold higher sensitivity of Deltapsi(m) to uncoupler. We conclude that the absence of the Surfl protein leads to the formation of incomplete COX complexes, 4 which in situ maintain rather high electron-transport activity, while their H+-pumping is impaired. Enzyme inactivation by the detergent in patient cells indicates instability of incomplete COX assemblies. (C) 2003 Elsevier B.V. All rights reserved.