Design of metabolic control for large flux changes

Metabolic Control Analysis has invalidated many traditional biochemical concepts of control, in particular the rate-limiting step. However, it has not been used to question the mechanisms by which pathway flux is thought to be controlled, such as the action of allosteric effectors or of covalent modification mechanisms. Here we use Control Analysis and computer simulation to examine the response of pathway segments to change in flux imposed by action on an enzyme outside the segment. Whether these segments contain near-equilibrium enzyme-catalysed reactions, cooperative enzymes, feedforward activation loops or feedback inhibition loops, their responses are significantly different from those observed in vivo. In particular, they do not exhibit the remarkable degrees of metabolite homoeostasis during large flux changes that have frequently been observed experimentally. On the other hand, near-constant levels of metabolites in spite of large changes of flux are consistent with our recent proposal that multi-site modulation--simultaneous activation of many pathway steps-is the normal method by which metabolism is controlled.