Extraction of mitochondrial proteins by volatile anesthetics

Heavy beef heart mitochondria were exposed to controlled concentrations of several volatile anesthetics including halothane, enflurane and chloroform. These anesthetics caused a concentration-dependent release of protein from mitochondria with maximal release occurring at 17.5% halothane and less release at lower and higher concentrations. The proteins released into the supernatants were analyzed by electrophoresis on slab gels containing a 6--20% gradient of acrylamide. The anesthetics caused the release of several polypeptides from mitochondria into the incubation medium; the major polypeptides released had molecular weights of 78 000; 48 000; 47 000; 43 000; 32 000 and 22 000. Two of these were identified by enzyme analysis and by co-electrophoresis with crystalline enzymes as the subunits of aspartate aminotransferase (43 000 daltons; EC 2.6.1.1) and malate dehydrogenase (32 000 daltons; EC 1.1.1.37). Mitochondria exposed to saturated halothane vapors were similar ultrastructurally to controls except that the halothane mitochondria appeared uncoupled. Similar results were obtained with O2 or N2 as carrier gas.