
==== Front
bioRxiv
BIORXIV
bioRxiv
2692-8205
Cold Spring Harbor Laboratory

10.1101/2024.09.05.611520
preprint
1
Article
Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
Skeens Erin http://orcid.org/0000-0002-0580-1291

Maschietto Federica http://orcid.org/0000-0002-5995-2765

Manjula Ramu http://orcid.org/0000-0002-2237-2599

Shillingford Shanelle http://orcid.org/0000-0003-2320-552X

Lolis Elias J. http://orcid.org/0000-0002-7902-7868

Batista Victor S.
Bennett Anton M. http://orcid.org/0000-0001-5187-7599

Lisi George P. http://orcid.org/0000-0001-8878-5655

05 9 2024
2024.09.05.611520https://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
http://biorxiv.org/lookup/doi/10.1101/2024.09.05.611520
nihpp-2024.09.05.611520.pdf
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 contributes to the interaction with p38 MAPK and JNK, thereby promoting dephosphorylation. Collectively, these results highlight the role of Y435 in the allosteric site as a novel mode of MKP5 regulation by p38 MAPK and JNK.
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pmc
