THE RECONSTITUTED CARNITINE CARRIER FROM RAT-LIVER MITOCHONDRIA - EVIDENCE FOR A TRANSPORT MECHANISM DIFFERENT FROM THAT OF THE OTHER MITOCHONDRIAL TRANSLOCATORS

The transport mechanism of the reconstituted carnitine carrier purified from rat liver mitochondria was investigated kinetically. The half-saturation constant (K-m) for carnitine on the internal side of the liposomal membrane (8.7 mM) was found to be much higher than that determined for the external surface (0.45 mM). The exclusive presence of a single transport affinity for carnitine on each side of the membrane indicated a unidirectional insertion of the carnitine carrier into the proteoliposomes, most probably right-side-out with respect to mitochondria. Under these defined conditions bisubstrate initial velocity studies of homologous (carnitine/carnitine) and heterologous (carnitine/acylcarnitine) antiport were performed by varying both the internal and external substrate concentrations. The kinetic patterns obtained showed that the ratio K-m/V-max is not influenced by the second (non-varied) substrate, which indicates a ping-pong mechanism. The carnitine carrier thus differs from all other mitochondrial carriers analyzed so far in the reconstituted state, for which a common sequential type of reaction mechanism has been found.