Membrane-bound fructose 1,6-bisphosphate aldolase: catalytic activity and mechanisms of desorption.

Fructose 1,6-bisphosphate aldolase [EC 4.1.2.13] in rat liver was found to be bound to the intracellular membraneous structures such as microsomes and nuclear membranes when the animals were fasted for 48 hr or administered tryptophan. Upon refeeding the rats the aldolase was released into the cytosol. The membrane-bound aldolase was almost inactive, showing about 50-fold larger Km and a smaller Vmax (37%) as compared with those of the free enzyme. The enzyme was released cooperatively from the membrane by exposure to fructose 1,6-bisphosphate, glyceraldehyde 3-phosphate or dihydroxyacetone phosphate at low concentrations. Apparent desorption constants (Kd, concentrations necessary for 50% desorption of enzyme) for fructose 1,6-bisphosphate of the enzymes bound to microsomes, mitochondria and nuclei were estimated to be 8 X 10(-5), 6.1 X 10(-6), and 4.8 X 10(-6)M, respectively, at pH 7.3. With the microsome-bound enzyme Kd values of 3.9 X 10(-4), 4.1 X 10(-4), 2.7 X 10(-3), 1.1 X 10(-2) and 2.0 X 10(-2) M were obtained for glyceraldehyde 3-phosphate, dihydroxyacetone phosphate, fructose 1-phosphate, fumarate, and KCl, respectively. Strong cooperativities were observed in the enzyme desorption by the substances which showed large Kd values.