Metabolic flux control analysis of branch points: an improved approach to obtain flux control coefficients from large perturbation data

An overview of published approaches for the metabolic flux control analysis of branch points revealed that often not all fundamental constraints on the flux control coefficients have been taken into account. This has led to contradictory statements in literature on the minimum number of large perturbation experiments required to estimate the complete set of flux control coefficients C-j for a metabolic branch point. An improved calculation procedure, based on approximate Lin-log reaction kinetics, is proposed, providing explicit analytical solutions of steady state fluxes and metabolite concentrations as a function of large changes in enzyme levels. The obtained solutions allow direct calculation of elasticity ratios from experimental data and subsequently all C-j-values from the unique relation between elasticity ratio's and flux control coefficients. This procedure ensures that the obtained C-j-values satisfy all fundamental constraints. From these it follows that for a three enzyme branch point only one characterised or two uncharacterised large flux perturbations are sufficient to obtain all C-j- values. The improved calculation procedure is illustrated with four experimental cases. (C) 2004 Elsevier Inc. All rights reserved.