Inbiochemistry, anEadie–Hofstee plot (orEadie–Hofstee diagram) is a graphical representation of theMichaelis–Menten equation inenzyme kinetics. It has been known by various different names, includingEadie plot,Hofstee plot andAugustinsson plot. Attribution toWoolf is often omitted, because althoughHaldane and Stern^[1] credited Woolf with the underlying equation, it was just one of the three linear transformations of the Michaelis–Menten equation that they initially introduced. However, Haldane indicated in 1957 that Woolf had indeed found the three linear forms:^[2]

In 1932, Dr. Kurt Stern published a German translation of my bookEnzymes, with numerous additions to the English text. On pp. 119–120, I described some graphical methods, stating that they were due to my friend Dr. Barnett Woolf. [...] Woolf pointed out that linear graphs are obtained whenv is plotted againstvx^-1,v^-1 againstx^-1, orv^-1x againstx, the first plot being most convenient unless inhibition is being studied.

The simplest equation for the rate${\displaystyle v}$ of an enzyme-catalysed reaction as a function of the substrate concentration${\displaystyle a}$ is the Michaelis-Menten equation, which can be written as follows:

${\displaystyle v=\frac{Va}{K_{\mathrm{m}}+a}}$

in which${\displaystyle V}$ is the rate at substrate saturation (when${\displaystyle a}$ approaches infinity, orlimiting rate, and${\displaystyle K_{\mathrm{m}}}$ is the value of${\displaystyle a}$ at half-saturation, i.e. for${\displaystyle v=0.5V}$, known as theMichaelis constant. Eadie^[3] and Hofstee^[4] transformed this intostraight-line relationship. Multiplication of both sides by${\displaystyle \frac{K_{\mathrm{m}}+a}{a}}$ gives:

${\displaystyle K_{\mathrm{m}}\cdot \frac{v}{a}+v=V}$

This can be directly rearranged to express a straight-line relationship:

${\displaystyle v=V-K_{\mathrm{m}}\cdot \frac{v}{a}}$

which shows that a plot of${\displaystyle v}$ against${\displaystyle v/a}$ is a straight line with intercept${\displaystyle V}$ on the ordinate, and slope${\displaystyle -K_{\mathrm{m}}}$ (Hofstee plot).

In the Eadie plot the axes are reversed:

${\displaystyle \frac{v}{a}=\frac{V}{K_{\mathrm{m}}}-\frac{1}{K_{\mathrm{m}}}\cdot v}$

with intercept$\frac{{\displaystyle V}}{K_{\mathrm{m}}}$ on the ordinate, and slope${\displaystyle -\frac{1}{K_{\mathrm{m}}}}$.

These plots are kinetic versions of theScatchard plot used in ligand-binding experiments.

The plot is occasionally attributed to Augustinsson^[5] and referred to theWoolf–Augustinsson–Hofstee plot^[6][7][8] or simply theAugustinsson plot.^[9] However, although Haldane, Woolf or Eadie were not explicitly cited when Augustinsson introduced the${\displaystyle v}$ versus${\displaystyle v/a}$ equation, both the work of Haldane^[10] and of Eadie^[3] are cited at other places of his work and are listed in his bibliography.^{[5]: 169 and 171}

Experimental error is usually assumed to affect the rate${\displaystyle v}$ and not the substrate concentration${\displaystyle a}$, so${\displaystyle v}$ is thedependent variable.^[11] As a result, both ordinate${\displaystyle v}$ and abscissa${\displaystyle v/a}$ are subject to experimental error, and so the deviations that occur due to error are not parallel with the ordinate axis but towards or away from the origin. As long as the plot is used forillustrating an analysis rather than forestimating the parameters, that matters very little. Regardless of these considerations various authors^[12][13][14] have compared the suitability of the various plots for displaying and analysing data.

Like other straight-line forms of the Michaelis–Menten equation, the Eadie–Hofstee plot was used historically for rapid evaluation of the parameters${\displaystyle K_{\mathrm{m}}}$ and${\displaystyle V}$, but has been largely superseded bynonlinear regression methods that are significantly more accurate when properly weighted and no longer computationally inaccessible.

As the ordinate scale spans the entire range of theoretically possible${\displaystyle v}$ vales, from${\displaystyle 0}$ to${\displaystyle V}$ one can see at a glance at an Eadie–Hofstee plot how well theexperimental design fills the theoretical design space, and the plot makes it impossible to hide poor design. By contrast, the other well known straight-line plots make it easy to choose scales that suggest that the design is better than it is. Faulty design, as shown in the right-hand diagram, is common with experiments with a substrate that is not soluble enough or too expensive to use concentrations above${\displaystyle K_{\mathrm{m}}}$, and in this case${\displaystyle V/K_{\mathrm{m}}}$ cannot be estimated satisfactorily. The opposite case, with${\displaystyle a}$ values concentrated above${\displaystyle K_{\mathrm{m}}}$ (left-hand diagram) is less common but not unknown, as for example in a study of nitrate reductase.^[15]

• Michaelis–Menten kinetics
• Lineweaver–Burk plot
• Hanes–Woolf plot
• Direct linear plot

• Diagrams
• Enzyme kinetics
• Biochemistry
• Biotechnology
• Eponyms in biology
• Eponymous diagrams of chemistry
• Molecular biology

• Articles with short description
• Short description matches Wikidata