Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase

Abstract Dual-specificity mitogen-activated protein kinase (MAPK) phosphatases (MKPs) directly dephosphorylate and inactivate the MAPKs. Although the catalytic mechanism of dephosphorylation of the MAPKs by the MKPs is established, a complete molecular picture of the regulatory interplay between the...

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Main Authors: Erin Skeens, Federica Maschietto, Ramu Manjula, Shanelle Shillingford, James Murphy, Elias J. Lolis, Victor S. Batista, Anton M. Bennett, George P. Lisi
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62150-w
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author Erin Skeens
Federica Maschietto
Ramu Manjula
Shanelle Shillingford
James Murphy
Elias J. Lolis
Victor S. Batista
Anton M. Bennett
George P. Lisi
author_facet Erin Skeens
Federica Maschietto
Ramu Manjula
Shanelle Shillingford
James Murphy
Elias J. Lolis
Victor S. Batista
Anton M. Bennett
George P. Lisi
author_sort Erin Skeens
collection DOAJ
description Abstract Dual-specificity mitogen-activated protein kinase (MAPK) phosphatases (MKPs) directly dephosphorylate and inactivate the MAPKs. Although the catalytic mechanism of dephosphorylation of the MAPKs by the MKPs is established, a complete molecular picture of the regulatory interplay between the MAPKs and MKPs still remains to be fully explored. Here, we sought to define the molecular mechanism of MKP5 regulation through an allosteric site within its catalytic domain. We demonstrate using crystallographic and NMR spectroscopy approaches that residue Y435 is required to maintain the structural integrity of the allosteric pocket. Along with molecular dynamics simulations, these data provide insight into how changes in the allosteric pocket propagate conformational flexibility in the surrounding loops to reorganize catalytically crucial residues in the active site. Furthermore, Y435 is required for the interaction with p38 MAPK and JNK, thereby promoting dephosphorylation. Collectively, these results demonstrate critical roles for the allosteric site in coordinating both MKP5 catalysis and MAPK binding.
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spelling doaj-art-67993377e7b84121a264dd698e8cb25f2025-08-20T04:03:03ZengNature PortfolioNature Communications2041-17232025-07-0116111410.1038/s41467-025-62150-wDynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphataseErin Skeens0Federica Maschietto1Ramu Manjula2Shanelle Shillingford3James Murphy4Elias J. Lolis5Victor S. Batista6Anton M. Bennett7George P. Lisi8Department of Molecular Biology, Cell Biology and Biochemistry, Brown UniversityDepartment of Chemistry, New York UniversityDepartment of Pharmacology, Yale School of Medicine, Yale University School of MedicineDepartment of Chemistry, Yale UniversityDepartment of Pharmacology, Yale School of Medicine, Yale University School of MedicineDepartment of Pharmacology, Yale School of Medicine, Yale University School of MedicineDepartment of Chemistry, Yale UniversityDepartment of Pharmacology, Yale School of Medicine, Yale University School of MedicineDepartment of Molecular Biology, Cell Biology and Biochemistry, Brown UniversityAbstract Dual-specificity mitogen-activated protein kinase (MAPK) phosphatases (MKPs) directly dephosphorylate and inactivate the MAPKs. Although the catalytic mechanism of dephosphorylation of the MAPKs by the MKPs is established, a complete molecular picture of the regulatory interplay between the MAPKs and MKPs still remains to be fully explored. Here, we sought to define the molecular mechanism of MKP5 regulation through an allosteric site within its catalytic domain. We demonstrate using crystallographic and NMR spectroscopy approaches that residue Y435 is required to maintain the structural integrity of the allosteric pocket. Along with molecular dynamics simulations, these data provide insight into how changes in the allosteric pocket propagate conformational flexibility in the surrounding loops to reorganize catalytically crucial residues in the active site. Furthermore, Y435 is required for the interaction with p38 MAPK and JNK, thereby promoting dephosphorylation. Collectively, these results demonstrate critical roles for the allosteric site in coordinating both MKP5 catalysis and MAPK binding.https://doi.org/10.1038/s41467-025-62150-w
spellingShingle Erin Skeens
Federica Maschietto
Ramu Manjula
Shanelle Shillingford
James Murphy
Elias J. Lolis
Victor S. Batista
Anton M. Bennett
George P. Lisi
Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
Nature Communications
title Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
title_full Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
title_fullStr Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
title_full_unstemmed Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
title_short Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
title_sort dynamic and structural insights into allosteric regulation on mkp5 a dual specificity phosphatase
url https://doi.org/10.1038/s41467-025-62150-w
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