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Detailed studies correlating changes in mitochondrial optical density, packed volume, and ultrastructure associated with osmotically-induced swelling were performed . Various swelling states were established by incubating mitochondria (isolated in 0 .25 M sucrose) at 0°C for 5 min in series of KCI and sucrose solutions ranging in tonicity from 250 to 3 milliosmols . Reversibility of swelling was determined by examining mitochondria exposed to 250 milliosmols media after they had been induced to swell . Swelling induced by lowering the ambient tonicity to approximately 130 (liver mitochondria) and 90 (heart mitochondria) milliosmols involves primarily swelling of the inner compartment within the intact outer membrane . Decreasing the ambient tonicity beyond this level results in rupture of the outer membrane and expansion of the inner compartment through the break . The maximum extent of swelling, corresponding with complete unfolding of the cristae and an increase in over-all mitochondrial volume of approximately 6-fold (liver mitochondria) and 11-fold (heart mitochondria), is reached at approximately 15 (liver mitochondria) and 3 (heart mitochondria) milliosmols . Exposure of liver mitochondria to media of lower tonicity results in irreversibility of inner compartment swelling and escape of matrix material . These changes appear to result from increased inner membrane permeability, possibly due to stretching .

The inner membranes of isolated bovine heart mitochondria undergo pronounced contraction upon being exposed to exogenous adenosine diphosphate (ADP), adenosine triphosphate (ATP), and certain other hlgh-energy phosphate compounds. Contraction results in decrease of inner membrane expanse which in turn results in decrease of intracristal space and increase of mitochondrial optical density (OD). The magnitude of the OD change appears to be proportional to the degree of contraction Half-maximal contraction can be achieved with ADP or ATP at concentrations as low as about 0 3 /zM. Atractyloside at concentrations as low as about 1.2 nmol/mg mitochondrial protein completely inhibits the contraction. It is concluded from these and other observations that inner membrane contraction occurs as a result of adenine nucleotide binding to the carrier involved in the exchange of adenine nucleotides across the inner mitochondrial membrane.

Both the rate and the driving force of a reaction can be expressed in terms of the concentrations of the reactants and products. Consequently, rate and driving force can be expressed as a function of each other. This has been done for a single-reactant, single-product, uncatalysed reaction and its enzyme-catalysed equivalent using the van't Hoff reaction isotherm and Haldane's generalized Michaelis-Menten rate equation, the primary objective being explanation of the exponential and sigmoidal relationships between reaction rate and delta mu H+ commonly observed in studies on chemiosmotic reactions. Acquisition of a purely thermodynamic rate vs. driving-force relationship requires recognition of the intensive and extensive variables and maintenance of the extensive variables constant. This relationship is identical for the two reactions and is hyperbolic or sigmoidal, depending on whether the equilibrium constant is smaller or larger than unity. In the case of the catalysed reaction, acquisition of the purely thermodynamic relationship requires the assumption that the enzyme be equally effective in catalysing the forward and backward reactions. If this condition is not met, the relationship is modified by the enzyme in a manner which can be determined from the ratio of the Michaelis constants of the reactant and product. Under conditions of enzyme saturation in respect to reactant+product, the rate vs. driving-force relationship is determined exclusively by the thermodynamics of the reaction and a single kinetic parameter, the magnitude of which is determined by the relative effectiveness of the enzyme in catalysing the forward and backward reactions. In view of this finding, it is pointed out that, since the catalytic components of chemiosmotic reactions appear to be saturated with respect to the reactant-product pair that is varied in experimental rate vs. delta mu H+ determinations, and that, since many complex enzymic reactions conform to the simple Michaelis-Menten equation with respect to a single reactant-product pair when the concentrations of all other reactants and products are maintained constant, one might expect to be capable of simulating the experimental relationships simply from knowledge of the thermodynamics of the reaction and the relative effectiveness of the catalytic component in catalysing the forward and backward reactions using the simple Michaelis-Menten equation. That this expectation appears to be largely correct is demonstrated with model reactions.(ABSTRACT TRUNCATED AT 400 WORDS)

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