Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon Plots in Non-steady-state Situations

Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon plots can only be used when a true initial rate is measured. Despite the fact that this point has often been stressed, it is far too often ignored in favour of restricting the assay time to one where low amounts of substrate are used. When one or several irreversible and slow steps occur with an inactivator during the incubation of a ternary enzyme-substrate-inactivator mixture, the rate of the enzyme-catalysed reaction progressively decreases. Even under these conditions, the present computer simulations investigations show that apparently linear Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon graphs can be obtained when the amount of product formed is mistakenly assumed to represent the true initial rate. Moreover, the observed pattern can change with time, going for instance from non-competitive to competitive. "Ki's" measured under these conditions also vary with time and bear little relationship to the true constants involved in the interaction.