Modulation of enzyme activity by inhibition and activation plays an important physiological role in regulation of cellular metabolism. Compared to the wealth of information available regarding inhibition of metabolic pathways, little is known about activation. Limited proteolysis of zymogens exemplifies irreversible activation. Reversible activation may involve post-translational modifications or dissociable binding of small molecules. Sometimes, chemical modification may also activate enzymes. The influence of small molecules on the reversible binding and activation of enzymes is summarized.