Measurement of enzyme activity.
To study and understand the nature of living cells, scientists have continually employed traditional biochemical techniques aimed to fractionate and characterize a designated network of macromolecular components required to carry out a particular cellular function. At the most rudimentary level, cellular functions ultimately entail rapid chemical transformations that otherwise would not occur in the physiological environment of the cell. The term enzyme is used to singularly designate a macromolecular gene product that specifically and greatly enhances the rate of a chemical transformation. Purification and characterization of individual and collective groups of enzymes has been and will remain essential toward advancement of the molecular biological sciences; and developing and utilizing enzyme reaction assays is central to this mission. First, basic kinetic principles are described for understanding chemical reaction rates and the catalytic effects of enzymes on such rates. Then, a number of methods are described for measuring enzyme-catalyzed reaction rates, which mainly differ with regard to techniques used to detect and quantify concentration changes of given reactants or products. Finally, short commentary is given toward formulation of reaction mixtures used to measure enzyme activity. Whereas a comprehensive treatment of enzymatic reaction assays is not within the scope of this chapter, the very core principles that are presented should enable new researchers to better understand the logic and utility of any given enzymatic assay that becomes of interest.
Harris
T K
TK
Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, Florida, USA.
Keshwani
M M
MM
D000818:Animals; D002384:Catalysis; D057075:Enzyme Assays / Q000379:methods*; D004798:Enzymes / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical
D004798:Enzymes