Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding
Summary: Activation of myosin light chain kinase (MLCK) by calcium ions (Ca2+) and calmodulin (CaM) plays an important role in numerous cellular functions including vascular smooth muscle contraction and cellular motility. Despite extensive biochemical analysis, aspects of the mechanism of activatio...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-04-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004223002237 |
| _version_ | 1827987202283405312 |
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| author | Melissa J.S. MacEwen Domnita-Valeria Rusnac Henok Ermias Timothy M. Locke Hayden E. Gizinski Joseph P. Dexter Yasemin Sancak |
| author_facet | Melissa J.S. MacEwen Domnita-Valeria Rusnac Henok Ermias Timothy M. Locke Hayden E. Gizinski Joseph P. Dexter Yasemin Sancak |
| author_sort | Melissa J.S. MacEwen |
| collection | DOAJ |
| description | Summary: Activation of myosin light chain kinase (MLCK) by calcium ions (Ca2+) and calmodulin (CaM) plays an important role in numerous cellular functions including vascular smooth muscle contraction and cellular motility. Despite extensive biochemical analysis, aspects of the mechanism of activation remain controversial, and competing theoretical models have been proposed for the binding of Ca2+ and CaM to MLCK. The models are analytically solvable for an equilibrium steady state and give rise to distinct predictions that hold regardless of the numerical values assigned to parameters. These predictions form the basis of a recently proposed, multi-part experimental strategy for model discrimination. Here we implement this strategy by measuring CaM-MLCK binding using an in vitro FRET system. Interpretation of binding data in light of the mathematical models suggests a partially ordered mechanism for binding CaM to MLCK. Complementary data collected using orthogonal approaches that assess CaM-MLCK binding further support this conclusion. |
| first_indexed | 2024-04-09T23:43:10Z |
| format | Article |
| id | doaj.art-dfa9997666004496bd94070810bacbc7 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-04-09T23:43:10Z |
| publishDate | 2023-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-dfa9997666004496bd94070810bacbc72023-03-18T04:42:30ZengElsevieriScience2589-00422023-04-01264106146Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase bindingMelissa J.S. MacEwen0Domnita-Valeria Rusnac1Henok Ermias2Timothy M. Locke3Hayden E. Gizinski4Joseph P. Dexter5Yasemin Sancak6Department of Pharmacology, University of Washington, Seattle, WA 98195, USADepartment of Pharmacology, University of Washington, Seattle, WA 98195, USADepartment of Pharmacology, University of Washington, Seattle, WA 98195, USADepartment of Pharmacology, University of Washington, Seattle, WA 98195, USADepartment of Pharmacology, University of Washington, Seattle, WA 98195, USAData Science Initiative and Department of Human Evolutionary Biology, Harvard University, Cambridge, MA 02138, USA; Corresponding authorDepartment of Pharmacology, University of Washington, Seattle, WA 98195, USA; Corresponding authorSummary: Activation of myosin light chain kinase (MLCK) by calcium ions (Ca2+) and calmodulin (CaM) plays an important role in numerous cellular functions including vascular smooth muscle contraction and cellular motility. Despite extensive biochemical analysis, aspects of the mechanism of activation remain controversial, and competing theoretical models have been proposed for the binding of Ca2+ and CaM to MLCK. The models are analytically solvable for an equilibrium steady state and give rise to distinct predictions that hold regardless of the numerical values assigned to parameters. These predictions form the basis of a recently proposed, multi-part experimental strategy for model discrimination. Here we implement this strategy by measuring CaM-MLCK binding using an in vitro FRET system. Interpretation of binding data in light of the mathematical models suggests a partially ordered mechanism for binding CaM to MLCK. Complementary data collected using orthogonal approaches that assess CaM-MLCK binding further support this conclusion.http://www.sciencedirect.com/science/article/pii/S2589004223002237BiochemistryBiochemical mechanismIn silico biology |
| spellingShingle | Melissa J.S. MacEwen Domnita-Valeria Rusnac Henok Ermias Timothy M. Locke Hayden E. Gizinski Joseph P. Dexter Yasemin Sancak Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding iScience Biochemistry Biochemical mechanism In silico biology |
| title | Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding |
| title_full | Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding |
| title_fullStr | Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding |
| title_full_unstemmed | Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding |
| title_short | Mathematical modeling and biochemical analysis support partially ordered calmodulin-myosin light chain kinase binding |
| title_sort | mathematical modeling and biochemical analysis support partially ordered calmodulin myosin light chain kinase binding |
| topic | Biochemistry Biochemical mechanism In silico biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004223002237 |
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