<b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b>
<p dir="ltr">The Michaelis constant (<i>K</i><sub>m</sub>) is a key parameter characterizing enzymatic reactions; however, determining <i>K</i><sub>m</sub> is prone to significant inaccuracies, and no practical method currently exists t...
محفوظ في:
| المؤلف الرئيسي: | |
|---|---|
| مؤلفون آخرون: | , , , , , |
| منشور في: |
2025
|
| الموضوعات: | |
| الوسوم: |
إضافة وسم
لا توجد وسوم, كن أول من يضع وسما على هذه التسجيلة!
|
| الملخص: | <p dir="ltr">The Michaelis constant (<i>K</i><sub>m</sub>) is a key parameter characterizing enzymatic reactions; however, determining <i>K</i><sub>m</sub> is prone to significant inaccuracies, and no practical method currently exists to assess the accuracy of <i>K</i><sub>m</sub> quantitatively. Here, we introduce a method for quantitatively assessing <i>K</i><sub>m</sub> accuracy using the concept of the Accuracy Confidence Interval (ACI). The ACI defines a range within which the true value of a parameter determined by nonlinear regression lies and is calculated through a combination of error-propagation and regression-stability analyses for a given regression model. We recently developed a workflow to find the ACI of the equilibrium dissociation constant (<i>K</i><sub>d</sub>) of affinity complexes, utilizing a standard nonlinear regression model for fitting binding isotherms of complexes with 1:1 stoichiometry. In this work, we demonstrate for the first time, that this "binding-isotherm" model can also be applied to the determination of <i>K</i><sub>m</sub>, suggesting that the ACI of <i>K</i><sub>m</sub> can be computed with the same workflow upon simply renaming the terms. Our findings advocate for the use of the binding-isotherm model in <i>K</i><sub>m</sub> determination since this model works for the whole range of total enzyme concentration and it enables a quantitative assessment of the accuracy. As the ACI of <i>K</i><sub>m</sub> can be easily computed with a user-friendly web app (<a href="https://aci.sci.yorku.ca" target="_blank">https://aci.sci.yorku.ca</a>), we anticipate quick and widespread adoption of <i>K</i><sub>m</sub> accuracy assessment by the research community.</p> |
|---|