Subcellular distribution of enzymes determined by rapid digitonin fractionation of isolated hepatocytes.

摘要: Conditions were determined for rapid separation of cytosolic and mitochondrial compartments by digitonin fractionation rat hepatocytes. The minimum time required enzyme markers decreased rapidly with increasing temperature. Kyro EOB, a non-ionic detergent, increases the release enzymes, particularly at lower temperatures. Experimental procedures are described greater than 90% enzymes less 2% in 3s. By using appropriate concentrations EOB medium maintained 1°C exposure to medium, nearly separate patterns obtained cytosol, matrix intermembrane space. distribution that exist more one these was quantified comparing their rates those marker enzymes. cytosol/mitochondrial-matrix distributions such hepatocytes from starved rats 16%/84% aspartate aminotransferase, 34%/66% fumarase 77%/23% ATP citrate lyase. In induced synthesize lyase starvation re-feeding, ratio had increased 95%/5%. maximum cytosol/intermembrane-space adenylate kinase 8%/92%. A procedure is also treating commercial its solubility water about 1mg/ml 800mg/ml.