Quantitative proteomics: the copy number of pyruvate dehydrogenase is more than 102‐fold lower than that of complex III in human mitochondria

Pyruvate dehydrogenase (PDH) and complex III are two key protein complexes in mitochondrial metabolic activity. Using a novel quantitative Western blotting method, we find that PDH and complex III exist at a steady-state ratio of 1:100, 1:128 and 1:202 in HeLa cell extracts, fibroblast mitochondria and heart tissue mitochondria, respectively. This difference in stoichiometry is reflected in the immunogold labeling intensities of the two complexes and by the much more sparse distribution of PDH in fluorescence microscopy. In Rho0 fibroblasts there is a 64% reduction of complex III but the concentration of PDH remains the same as wild-type.