Kinetics of the alanine aminotransferase reaction in the mitochondrial and cell sap fractions of rat brain.
1. A reversible transamination reaction between L-glutamate and pyruvate, or L-alanine and 2-oxoglutarate, takes place in the mitochondrial and cell sap fractions of rat brain. 2. The maximum rate of the transamination reaction in both subfractions was observed in the presence of a keto- substrate concentration of 2.5 mM only, but an amino- donor concentration of 20 mM. 3. The apparent Menten-Michaelis constants for pyruvate and 2-oxoglutarate were of a 10(-4) M and for L-glutamate and L-alanine of a 10(-3) M order and were approximately the same for both fractions. 4. The ratio of the initial rate of the L-alanine + 2-oxoglutarate to the L-glutamate + pyruvate transamination reaction in the cell sap and mitochondrial fractions amounted to up to 2. 5. The apparent equilibrium constant derived from the Haldane equation was 7.01 for cell sap alanine aminotransferase and 4 for the mitochondrial enzyme. 6. Increasing pyridoxal-5'-phosphate concentrations in the incubation medium were accompanied by only non-significant stimulation of alanine aminotransferase activity in the mitochondrial and cell sap fractions. 7. A comparison of the kinetic data obtained on mitochondrial and cell sap alanine aminotransferases in vitro with the actual substrate concentrations in the transamination reaction in nervous tissue in vivo indicates that the direction of the transamination reaction in situ seems to be determined simply by compartmentation and by dynamic changes in amino- and keto- substrates in the mitochondrial and cell sap spaces.
D000410:Alanine Transaminase / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D003593:Cytoplasm / Q000201:enzymology* / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D066298:In Vitro Techniques; D007656:Ketoglutaric Acids / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D011773:Pyruvates / Q000378:metabolism; D051381:Rats
D005971:Glutamates; D007656:Ketoglutaric Acids; D011773:Pyruvates; D011732:Pyridoxal Phosphate; D000410:Alanine Transaminase