Hanes–Woolf plot

Hanes plot of a/v against a for Michaelis–Menten kinetics

In biochemistry, a Hanes–Woolf plot, Hanes plot, or plot of $a/v$ against $a$ is a graphical representation of enzyme kinetics in which the ratio of the initial substrate concentration $a$ to the reaction velocity $v$ is plotted against $a$ . It is based on the rearrangement of the Michaelis–Menten equation shown below:

{a \over v}={a \over V}+{K_{\mathrm {m} } \over V}

where $K_{\mathrm {m} }$ is the Michaelis constant and $V$ is the limiting rate.^[1]

J. B. S. Haldane stated, reiterating what he and K. G. Stern had written in their book,^[2] that this rearrangement was due to Barnet Woolf.^[3] However, it was just one of three transformations introduced by Woolf. It was first published by C. S. Hanes, though he did not use it as a plot.^[4] Hanes noted that the use of linear regression to determine kinetic parameters from this type of linear transformation generates the best fit between observed and calculated values of $1/v$ , rather than $v$ .^[4]^: 1415

Starting from the Michaelis–Menten equation:

v={{Va} \over {K_{\mathrm {m} }+a}}

we can take reciprocals of both sides of the equation to obtain the equation underlying the Lineweaver–Burk plot:

{1 \over v}={1 \over V}+{K_{\mathrm {m} } \over V}\cdot {1 \over a}

which can be multiplied on both sides by ${a}$ to give

{a \over v}={1 \over V}\cdot a+{K_{\mathrm {m} } \over V}

Thus in the absence of experimental error data a plot of ${a/v}$ against ${a}$ yields a straight line of slope $1/V$ , an intercept on the ordinate of ${K_{\mathrm {m} }/V}$ and an intercept on the abscissa of $-K_{\mathrm {m} }$ .

Like other techniques that linearize the Michaelis–Menten equation, the Hanes–Woolf plot was used historically for rapid determination of the kinetic parameters $K_{\mathrm {m} }$ , $V$ and $K_{\mathrm {m} }/V$ , but it has been largely superseded by nonlinear regression methods that are significantly more accurate and no longer computationally inaccessible. It remains useful, however, as a means to present data graphically.

References

^ The term maximum rate is often used, but not recommended by the IUBMB; see Cornish-Bowden, A (2014). "Current IUBMB recommendations on enzyme nomenclature and kinetics". Persp. Sci. 1: 74–87. doi:10.1016/j.pisc.2014.02.006.
^ Haldane, John Burdon Sanderson; Stern, Kurt Günter (1932). Allgemeine Chemie der Enzyme. Wissenschaftliche Forschungsberichte, Naturwissenschaftliche Reihe, herausgegeben von Dr. Raphael Eduard Liesegang. Vol. 28. Dresden and Leipzig: Theodor Steinkopff. pp. 119–120. OCLC 964209806.
^ Haldane, John Burdon Sanderson (1957). "Graphical methods in enzyme chemistry". Nature. 179 (4564): 832–832. Bibcode:1957Natur.179R.832H. doi:10.1038/179832b0. S2CID 4162570.
^ ^a ^b Hanes, Charles Samuel (1932). "Studies on plant amylases: The effect of starch concentration upon the velocity of hydrolysis by the amylase of germinated barley". Biochemical Journal. 26 (5): 1406–1421. doi:10.1042/bj0261406. PMC 1261052. PMID 16744959.

v t e Enzymes
Activity	Active site Binding site Catalytic triad Oxyanion hole Enzyme promiscuity Diffusion-limited enzyme Cofactor Enzyme catalysis
Regulation	Allosteric regulation Cooperativity Enzyme inhibitor Enzyme activator
Classification	EC number Enzyme superfamily Enzyme family List of enzymes
Kinetics	Enzyme kinetics Eadie–Hofstee diagram Hanes–Woolf plot Lineweaver–Burk plot Michaelis–Menten kinetics
Types	EC1 Oxidoreductases (list) EC2 Transferases (list) EC3 Hydrolases (list) EC4 Lyases (list) EC5 Isomerases (list) EC6 Ligases (list) EC7 Translocases (list)

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