Large-scale preparation of cytosolic and mitochondrial asparatate aminotransferases from pig heart.
A procedure is described for the large-scale preparation of the cytosolic and mitochondrial isoenzymes of aspartate aminotransferase from pig heart. The procedure consists of: 1. extraction of both isoenzymes by heat treatment of homogenates prepared from minced and frozen heat muscle; 2. separation of each isoenzyme on a hydroxyapatite column; 3. purification of each isoenzyme by combinations of heat treatment, ammonium sulfate fractionation and chromatography on ion-exchange cellulose columns. Purified preparations of each isoenzyme thus obtained were homogeneous proteins as judged from their spectral properties and behavior on polyacrylamide gel electrophoresis. Using the present procedure, 1.2 g of the cytosolic isoenzyme and 1.7 g of the mitochondrial isoenzyme were obtained from 20 kg of minced heart muscle.
D000645:Ammonium Sulfate; D000818:Animals; D001219:Aspartate Aminotransferases / Q000302:isolation & purification*; D005591:Chemical Fractionation / Q000379:methods; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D003600:Cytosol / Q000201:enzymology; D005374:Filtration / Q000379:methods; D006358:Hot Temperature; D006882:Hydroxyapatites; D007527:Isoenzymes / Q000302:isolation & purification*; D008929:Mitochondria, Heart / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology*; D013552:Swine
D006882:Hydroxyapatites; D007527:Isoenzymes; D001219:Aspartate Aminotransferases; D000645:Ammonium Sulfate