Human diseases with defects in oxidative phosphorylation 1. Decreased amounts of assembled oxidative phosphorylation complexes in mitochondrial encephalomyopathies

encephalomyopath y patients has been studied by two-dimensional electrophoresis (blue native PAGE/Tricine-SDS -PAGE). Only 20 mg muscle was required to identify and analyse complexes I, III, IV, and V after Coomassie staining. In most cases reduced amounts of the involved complex(es) correlated well with decreased enzyme activities. The reliability of the method was reflected by the constant mutual ratio of the complexes found in all controls. Deviations from normal ratios were found to be more sensitive indicators for a defect than the absolute quantities, which varied considerably within the control group both in the enzymic and in the electropho­ retic analysis. The effect of the mitochondrial tRNAL cu(UUR) mutation in mitochondrial myopathy, encepha­ lopathy, lactic acidosis, and stroke-like episodes on the amount of oxidative phosphorylation complexes was demonstrated for the first time directly on the protein level. In patients without known DNA mutations, specific defects of single complexes were identified. The new technique is a sensitive method for the identification of oxidative phosphorylation defects, complementary to enzymic measurements.