Concentration and partitioning of intermediates in the fructose bisphosphate aldolase reaction. Comparison of the muscle and liver enzymes.
Chemical analysis of enzyme reaction intermediates has been used to compare the liver and muscle isozymes of rabbit aldolase at equilibrium and in their steady states to determine if they have properties that favor the direction of flow of glycolytic intermediates in their tissues of origin. For both enzymes at saturating concentrations of fructose 1,6-P2, the sum of intermediates in the steady state agreed with the total active enzyme calculated to be present. The two half-reactions, characterized by fructose 1,6-bisphosphate(Fru-P2):aldehyde exchange and DHAP:proton exchange were found to be of different importance in determining the rate of reaction with Fru-P2 with the liver enzyme being much more limited in the processing of DHAP. The chemical interconversions within each half-reaction are generally rapid compared with the release of products. The greater sensitivity of liver aldolase to inhibition by aldehydes in Fru-P2 cleavage seems to be a normal consequence of the higher level of the eneamine of DHAP in the forward steady state with the liver enzyme and probably should not be ascribed to a greater intrinsic affinity. An earlier report (Grazi, E., and Trombetta, G. (1979) Eur. J. Biochem. 100, 197-202) purporting to show a special interaction of glyceraldehyde-3-P with liver enzyme prior to proton abstraction from DHAP could not be reproduced. Examples are presented from the data that validate the use of the analytical methods used for analysis of intermediates in the case of the Schiff's base aldolases.
D000818:Animals; D004099:Dihydroxyacetone Phosphate / Q000378:metabolism; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D009132:Muscles / Q000201:enzymology*; D009928:Organ Specificity; D010761:Phosphorus Radioisotopes; D011485:Protein Binding; D011817:Rabbits
D010761:Phosphorus Radioisotopes; D004099:Dihydroxyacetone Phosphate; D005634:Fructose-Bisphosphate Aldolase