In enzyme kinetics, a secondary plot uses the intercept or slope from several Lineweaver–Burk plots to find additional kinetic constants. For example, when a set of v by [S] curves from an enzyme with a ping–pong mechanism (varying substrate A, fixed substrate B) are plotted in a Lineweaver–Burk plot, a set of parallel lines will be produced. The following Michaelis–Menten equation relates the initial reaction rate v0 to the substrate concentrations [A] and [B]: The y-intercept of this equation is equal to the following: The y-intercept is determined at several different fixed concentrations of substrate B (and varying substrate A). The y-intercept values are then plotted versus 1/[B] to determine the Michaelis constant for substrate B, K_M^B, as shown in the Figure to the right. The slope is equal to K_M^B divided by v_\max and the intercept is equal to 1 over v_\max.

Secondary plot in inhibition studies

A secondary plot may also be used to find a specific inhibition constant, KI. For a competitive enzyme inhibitor, the apparent Michaelis constant is equal to the following: The slope of the Lineweaver-Burk plot is therefore equal to: If one creates a secondary plot consisting of the slope values from several Lineweaver-Burk plots of varying inhibitor concentration [I], the competitive inhbition constant may be found. The slope of the secondary plot divided by the intercept is equal to 1/KI. This method allows one to find the KI constant, even when the Michaelis constant and vmax values are not known.