A permeability barrier in mitochondria in ;high-energy states' against positively charged disulphides.

1. In rat liver mitochondria in state 1 or 4 there is a permeability barrier against cystamine, probably in the inner membrane. 2. The permeability barrier was broken (a) when oxidative phosphorylation was uncoupled, (b) when the respiratory chain was inhibited or in anaerobiosis, or (c) when phosphate was added in the absence of exogenous substrate. Under these conditions increased amounts of [(35)S]cystamine residues were bound to matrix proteins. 3. It appears that the permeability barrier against cystamine in mitochondria reflects a ;high-energy state'. A gradual increase in the permeability for cystamine strikingly coincided with the loss of respiratory control induced by increasing concentrations of different uncoupling agents. 4. Cystamine caused uncoupling of oxidative phosphorylation in state 2 or 5, but not in state 1, 3 or 4. The uncoupling effect of cystamine was dependent on the phosphorylation potential. ATP counteracted, whereas ADP potentiated, the uncoupling by cystamine. 5. The variable penetration of cystamine appears to depend on its positive charge, since a dication derivative, NNN'N'-tetramethylcystamine, has a similar pattern of penetration, whereas an uncharged derivative, NN'-diacetylcystamine, penetrates rapidly into mitochondria irrespective of their metabolic state. 6. It is suggested that a charge barrier is present in or across the inner mitochondrial membrane in ;high-energy states'.