Study of interaction kinetics and enzymatic pathways

Determination of the transient kinetics of protein-ligand interactions and of enzymatic processes

Transient kinetics techniques (stopped-flow, quench-flow, cryoenzymology) are used to measure enzymatic kinetics on the millisecond time scale. The main applications of these techniques are :

• Protein-ligand interaction measurements: k_on, k_off, K_d.
• Determination of reaction mechanisms: rate constants of each step, determination of the kinetically limiting step.
• Characterization of enzymatic inhibitors: type of inhibition, K_i.

Example of transient kinetics obtained with quench-flow and relationship between the transient phases and different steps of the reaction pathway.

Stopped-flow

Quench-flow

Cryo Quench-flow