Kinetic and binding properties of the oxoglutarate translocator of rat-heart mitochondria.

The kinetic study of the oxoglutarateout/malatein exchange through the inner mitochondrial membrane of rat-heart mitochondria has been compelted and extended to higher external-oxoglutarate and to lower internal-malate concentrations. It has been found that the external oxoglutarate inhibits the exchange at high concentration. This excess-substrate inhibition is preceded by four jumps. The kinetic-saturation curve by the internal malate presents an apparent positive cooperativity that may be interpreted in different ways. The independence of the effects of the two substrates on the initial rate has been observed again and supports the conclusions reached in previous work. A method for the determination of oxoglutarate binding to the external face of the inner membrane is described. The binding curve shows four intermediary plateau regions that reflect significant apparent K-effects, alternatively negative and positive. For external-oxoglutarate concentrations below the region of excess-substrate inhibition, the binding-saturation curve and the kinetic-saturation curve are similar, demonstrating that K-effects are predominant. A particularly wide intermediary plateau that seems to correspond to half saturation of the active sites is common to both saturation curves. A clear lack of proportionality between the two curves at low oxoglutarate concentrations seems to indicate that more than one catalytic-rate constant is implied in the exchange kinetics. Two models of the oxoglutarate carrier are presented. Both lead to a minimum degree of 10:10 for the equation of the binding of oxoglutarate to the catalytic sites. In the first model this corresponds to ten subunits associated into a single oligomer while in the second model this results from a mixture of monomeric, dimeric, trimeric and tetrameric associations.