Systems analysis of the tricarboxylic acid cycle in Dictyostelium discoideum. I. The basis for model construction.

A steady-state model of the tricarboxylic acid cycle was constructed using a dynamic systems analysis computer program, METASIM. The model was based on radioactive tracer analyses which provided flux relationships and compartmented metabolite concentrations. Ten of the enzymes modeled were purified and characterized from Dictyostelium discoideum. Although experimentally determined enzyme mechanisms and constants were used in the model, Vmax values were found to be unreliable, i.e. they did not reflect enzyme activity in vivo. This value was therefore calculated as the only unknown in each enzyme kinetic equation and called Vvivo, to distinguish it from Vmax determined in vitro.