Problems in the experimental determination of substrate-specific H+/O ratios during respiration

Krabet al. (1984) have recently tried to resolve the long-standing controversy as to whether the mechanistic H⁺/O coupling ratio for electrons passing through sites II and III of the mammalian electron transport chain to O₂ is 6 or 8. Using a mathematical model they concluded that the higher number reported by Costaet al. (1984) was an overestimate because of the unaccounted for delayed response of the O₂ electrode. Responding to criticisms of Lehningeret al. (1985), they have recently used (Krab and Wikström, 1986) an improved mathematical model which shows that the higher number found by Costaet al. was probably due to an inadequate accounting for the effects of the proton leak process which accompanies the translocation process. The impression is left that the situation is now resolved in favor of the lower number. We agree that the procedures of Costaet al. do not properly account for the leak process, and provide further evidence in this paper of the magnitude of the problem. However, we disagree that the number 6.0, favored by Wikströmet al., rests on any more solid experimental support. We provide evidence here for this conclusion and raise the question as to whether or not any unique, fixed, integral number exists for the H⁺/O ratio accompanying the oxidation of a particular substrate.