Regulation of enzyme activity in the cell: effect of enzyme concentration.

The rapid development in our understanding of the regulation of enzyme activity makes it a high priority to ascertain whether the behavior of purified enzymes reflects their functional characteristics in vivo. Enzyme concentration is usually the most significant difference between routine in vitro assays and in vivo conditions, as it is well known that many intracellular enzymes are present in vivo at much higher concentrations than used in vitro. Various procedures are suitable for kinetic analysis at physiological concentrations of enzyme. Those more frequently used have been cell permeabilization, the utilization of purified enzymes at concentrations close to the in vivo range, and the addition of polyethylene glycol to increase the local protein concentration. In this review we briefly summarize observations on enzymes reported to exhibit concentration-dependent activity. The effect of enzyme concentration has been most thoroughly investigated in the case of phosphofructokinase. These studies may provide insight into the regulation of this important enzyme in the cell. The implications of both homologous and heterologous protein-protein interactions for the effect of enzyme concentration and their roles in the control of enzyme activity in vivo are also discussed.