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Use of modified double-reciprocal plot for enzyme kinetic parameters. A pitfall

N G Karanth, A K Srivastava

    Biochimica Et Biophysica Acta
    |September 9, 1980
    PubMed
    Summary

    The Lee and Wilson method for determining enzyme kinetics parameters V and Km can lead to significant errors, even with partial substrate consumption. Researchers recommend using the complete integrated Michaelis-Menten equation for accurate enzyme kinetic analysis.

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    Area of Science:

    • Biochemistry
    • Enzyme Kinetics

    Background:

    • The modified double-reciprocal plot by Lee and Wilson is a widely accepted method for enzyme kinetic parameter determination.
    • This method is believed to be accurate even when a significant portion of the substrate is consumed.

    Purpose of the Study:

    • To analytically evaluate the accuracy of the Lee and Wilson method for enzyme kinetic parameter determination.
    • To identify potential sources of error in the application of the modified double-reciprocal plot.

    Main Methods:

    • Analytical proof was used to assess the errors associated with the Lee and Wilson method.
    • Comparison of the modified double-reciprocal plot with the integrated Michaelis-Menten equation.

    Main Results:

    • The Lee and Wilson method can introduce substantial errors in enzyme kinetic parameter (V and Km) determination.

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  • Errors are significantly higher than previously assumed, particularly with substantial substrate consumption.

    • Conclusions:

    • The modified double-reciprocal plot by Lee and Wilson is not as accurate as previously thought.
    • The complete integrated Michaelis-Menten equation should be employed for precise determination of enzyme kinetic parameters.