Turnover of cytoplasmic and mitochondrial aspartate aminotransferase isozymes in mouse liver and transplantable hepatomas.
Activities of the cytoplasmic and mitochondrial isozymes of aspartate aminotransferase (aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1, AAT) in transplantable mouse hepatomas BW7756 and H-4 are reduced when compared to normal adult liver. Both proteins have been purified to homogeneity from a single preparation of mouse liver and monospecific antibodies raised to each isozyme. By quantitative immunotitration analysis, the activity of each isozyme in liver and hepatoma has been shown to correlate with levels of immunoprecipitable protein. Furthermore, for each isozyme, the liver versus hepatoma species is indistinguishable by heat inactivation kinetics, Km's for substrates, and molecular weights. Thus, the reduction of mitochondrial and cytoplasmic AAT activities in hepatoma tissue is due not to alterations in the catalytic activity of the enzyme molecules, but to a decrease in the number of enzyme molecules present. Turnover of the isozymes was studied in liver and hepatoma tissue using in vivo radiolabeling and specific immunoprecipitation techniques. The cytoplasmic isozyme has a similar rate of degradation in liver and hepatoma, while the rate of synthesis of this isozyme in hepatoma is approximately tenfold less than in liver. The mitochondrial isozyme is also degraded at a similar rate in both tissues, but the rate of synthesis is sixfold greater in normal liver tissue than in hepatoma. It is concluded that decreased amounts of both isozymes in hepatoma as compared to liver are the result of a reduction in the rate of synthesis of each isozyme without any change in the rate of degradation.
D000818:Animals; D001219:Aspartate Aminotransferases / Q000276:immunology / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D051379:Mice; D008930:Mitochondria, Liver / Q000201:enzymology*; D009368:Neoplasm Transplantation
D007527:Isoenzymes; D001219:Aspartate Aminotransferases