Cryoenzymology: Enzyme Action in Slow Motion

Knowledge of the existence and structure of intermediates on the reaction pathway is necessary before specific details of the mechanism may be successfully resolved. However, enzymatic catalysis is an extremely fast process. This rapidity of enzyme-catalyzed reactions and the short life times of intermediates represent a major problem in studying the dynamic processes which occur during catalysis, as they prevent the accumulation of intermediates under normal conditions for concentrations and time periods required by most high-resolution structural methods. Therefore, a method that would utilize specific substrates but would permit the detection and characterization of intermediates was highly desired. As one of the approaches to overcome this problem the use of cryoenzymology to allow the accumulation and stabilization of intermediates at very low temperatures was proposed. This review describes the contribution of Prof. Anthony L. Fink to cryoenzymology and shows how his work shaped this exciting area.