Application of a normalised plot to the study of ter ter enzyme systems

The kinetic characterisation of multisubstrate systems is not a trivial task. Common approaches simplify the experimental procedures by sequentially fixing saturating concentrations of different substrates/products, thereby attempting to isolate the influence of the varying molecule. Even after such tedious work, only apparent K m values can be determined, preventing serious comparison among differential substrate behaviours. Moreover, the choice among rival kinetic models is not statistically guaranteed; instead, classical tools such as re-plots continue to be used. Here, we report the application of a normalisation of kinetic data, formerly applied to simpler systems, to the description of ter ter systems. This data treatment is able to provide true K m values and a reliable description of the system, at the same time reducing the experimental work and statistically supporting the choice of kinetic schemes.