Using Isothermal Titration Calorimetry to Characterize Enzyme Kinetics

Enzymes are proteins that function as biological catalysts, which play crucial roles in the biochemical processes that occur in living organisms.  Understanding how enzymes function, and how to activate or inhibit their activity, is a core research focus for biochemists. Since many drug targets are enzymes, the development of new therapies requires understanding of how the target enzyme binds and catalyzes its natural substrate. Several drugs are therapeutic enzymes, which are injected into the patient to treat genetic disorders characterized by missing or defective enzymes. Enzymes are also important in other industries including food science, biofuels, and detergents.

Although enzymes were discovered in the mid-1800s, isolated and purified since the early 20th century, and cloned and expressed in recombinant systems since the 1970s, there is a continued need for efficient and detailed enzyme analysis to exploit the potential of enzyme-driven catalytic reactions. New enzymes are also being discovered. Reliable enzyme kinetics data are crucial to understand and control enzyme effectiveness and create next generation drugs.

Isothermal titration calorimetry (ITC) techniques have been successfully applied to study enzyme kinetics and inhibition. ITC is a well-established, versatile technique that is widely used for measuring reaction thermodynamics.  In this review, we discuss how ITC generates real-time, enzyme kinetics data, comparable to other enzyme assays.  In part 2, we discuss how to perform enzyme kinetics experiments with a MicroCal ITC system.

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