<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...
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2025
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| _version_ | 1852018188876775424 |
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| author | TONG YE WANG (12959627) |
| author2 | Parmeetpal Dhillon (19557283) Svetlana Krylova (6644106) Sebastian Schreiber (18523618) Dasantila Golemi-Kotra (19557287) Prof. Joachim Jose (19354272) Sergey Krylov (9416975) |
| author2_role | author author author author author author |
| author_facet | TONG YE WANG (12959627) Parmeetpal Dhillon (19557283) Svetlana Krylova (6644106) Sebastian Schreiber (18523618) Dasantila Golemi-Kotra (19557287) Prof. Joachim Jose (19354272) Sergey Krylov (9416975) |
| author_role | author |
| dc.creator.none.fl_str_mv | TONG YE WANG (12959627) Parmeetpal Dhillon (19557283) Svetlana Krylova (6644106) Sebastian Schreiber (18523618) Dasantila Golemi-Kotra (19557287) Prof. Joachim Jose (19354272) Sergey Krylov (9416975) |
| dc.date.none.fl_str_mv | 2025-07-25T22:25:14Z |
| dc.identifier.none.fl_str_mv | 10.6084/m9.figshare.26961733.v2 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/figure/_b_Introducing_Quantitative_Assessment_of_Michaelis_Constant_b_b_i_K_i_b_sub_strong_M_strong_sub_b_Accuracy_b_/26961733 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Catalysis and mechanisms of reactions Reaction kinetics and dynamics michaelismenten Accuracy of equilibrium constants Accuracy confidence interval (ACI) of parameters computed through nonlinear regression Precision Confidence Interval Regression stability error propagation algorithm enzymatic reaction path |
| dc.title.none.fl_str_mv | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| dc.type.none.fl_str_mv | Image Figure info:eu-repo/semantics/publishedVersion image |
| description | <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> |
| eu_rights_str_mv | openAccess |
| id | Manara_de22927f12f5deaa61d78fa9dbf2be95 |
| identifier_str_mv | 10.6084/m9.figshare.26961733.v2 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/26961733 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b>TONG YE WANG (12959627)Parmeetpal Dhillon (19557283)Svetlana Krylova (6644106)Sebastian Schreiber (18523618)Dasantila Golemi-Kotra (19557287)Prof. Joachim Jose (19354272)Sergey Krylov (9416975)Catalysis and mechanisms of reactionsReaction kinetics and dynamicsmichaelismentenAccuracy of equilibrium constantsAccuracy confidence interval (ACI) of parameters computed through nonlinear regressionPrecision Confidence IntervalRegression stabilityerror propagation algorithmenzymatic reaction path<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>2025-07-25T22:25:14ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.6084/m9.figshare.26961733.v2https://figshare.com/articles/figure/_b_Introducing_Quantitative_Assessment_of_Michaelis_Constant_b_b_i_K_i_b_sub_strong_M_strong_sub_b_Accuracy_b_/26961733CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/269617332025-07-25T22:25:14Z |
| spellingShingle | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> TONG YE WANG (12959627) Catalysis and mechanisms of reactions Reaction kinetics and dynamics michaelismenten Accuracy of equilibrium constants Accuracy confidence interval (ACI) of parameters computed through nonlinear regression Precision Confidence Interval Regression stability error propagation algorithm enzymatic reaction path |
| status_str | publishedVersion |
| title | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| title_full | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| title_fullStr | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| title_full_unstemmed | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| title_short | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| title_sort | <b>Introducing Quantitative Assessment of Michaelis Constant (</b><b><i>K</i></b><sub><strong>M</strong></sub><b>) Accuracy</b> |
| topic | Catalysis and mechanisms of reactions Reaction kinetics and dynamics michaelismenten Accuracy of equilibrium constants Accuracy confidence interval (ACI) of parameters computed through nonlinear regression Precision Confidence Interval Regression stability error propagation algorithm enzymatic reaction path |