Activation loop phosphorylation tunes conformational dynamics underlying Pyk2 tyrosine kinase activation

Zanela, Tânia Marina Palhano; Woudenberg, Alexzandrea; Bello, Karen G Romero; Underbakke, Eric S.

doi:10.1016/j.str.2023.02.003

Cited by 4 publications

(11 citation statements)

References 42 publications

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“…9,10 Indeed, previous studies established that disruption of the Pyk2 FERM F2:kinase C-lobe interface leads to increased autophosphorylation. 15 Hence, we sought to test whether the K60P mutation disrupts the global autoinhibitory conformation of Pyk2. We employed global hydrogen/deuterium exchange mass spectrometry (HDX-MS) to monitor the general uptake of deuterium throughout the intact protein.…”

Section: Resultsmentioning

confidence: 99%

“…The canonical mechanism for full activation of autophosphorylated Pyk2 involves Src docking and subsequent activation loop phosphorylation at Pyk2 residues Y579 and Y580. 14,15,26 Given the pronounced increase in Pyk2 autophosphorylation upon disruption of the putative FERM:linker interface, we tested whether a K60 variant also impacted the phosphorylation status of the activation loop. We monitored site-specific phosphorylation in WT and K60P Pyk2 FERM-kinase.…”

Section: Conformational Context Modulates Pyk2 Autophosphorylation Vi...mentioning

confidence: 99%

“…Although the mechanistic details of Pyk2 activation are obscured by limited high-resolution structural models, investigations dissecting Pyk2 regulation revealed key features. [13][14][15] Pyk2 autophosphorylates residue Y402 independently of Src kinase activity. Autophosphorylation is sufficient for Src to dock and outcompete the dynamic FERM-kinase autoinhibitory interface to phosphorylate the Pyk2 activation loop tyrosines (Y579, Y580).…”

Section: Introductionmentioning

confidence: 99%

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Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Zanela,

Zangiabadi,

Zhao

et al. 2024

RSC Chem. Biol.

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Section: Resultsmentioning

confidence: 99%

Section: Conformational Context Modulates Pyk2 Autophosphorylation Vi...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Zanela,

Zangiabadi,

Zhao

et al. 2024

RSC Chem. Biol.

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“…2a) 9,10 . Indeed, previous studies established that disruption of the Pyk2 FERM F2:kinase C-lobe interface leads to increased autophosphorylation 15 . Hence, we sought to test whether the K60P mutation disrupts the global autoinhibitory conformation of Pyk2.…”

Section: Conformational Context Modulates Pyk2 Autophosphorylation Vi...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Zanela,

Zangiabadi,

Zhao

et al. 2023

Preprint

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Pyk2 is a multi-domain non-receptor tyrosine kinase that serves dual roles as signaling enzyme and scaffold. Pyk2 activation involves a multi-stage cascade of conformational rearrangements and protein interactions initiated by autophosphorylation of a linker site. Linker phosphorylation recruits Src kinase, and Src-mediated phosphorylation of the Pyk2 activation loop confers full activation. The regulated accessibility of the initial Pyk2 autophosphorylation site remains unclear. We employed peptide-binding molecularly imprinted nanoparticles (MINPs) to probe the regulatory conformations controlling Pyk2 activation. MINPs differentiating local structure and phosphorylation state revealed that the Pyk2 autophosphorylation site is protected in the autoinhibited state. Activity profiling of Pyk2 variants implicated FERM and linker residues responsible for constraining the autophosphorylation site. MINPs targeting each Src docking site disrupt the higher-order kinase interactions critical for activation complex maturation. Ultimately, MINPs targeting key regulatory motifs establish a useful toolkit for probing successive activational stages in the higher-order Pyk2 activation complex.

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Regulation and signaling of the LIM domain kinases

Casanova‐Sepúlveda,

Boggon

2024

BioEssays

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The LIM domain kinases (LIMKs) are important actin cytoskeleton regulators. These proteins, LIMK1 and LIMK2, are nodes downstream of Rho GTPases and are the key enzymes that phosphorylate cofilin/actin depolymerization factors to regulate filament severing. They therefore perform an essential role in cascades that control actin depolymerization. Signaling of the LIMKs is carefully regulated by numerous inter‐ and intra‐molecular mechanisms. In this review, we discuss recent findings that improve the understanding of LIM domain kinase regulation mechanisms. We also provide an up‐to‐date review of the role of the LIM domain kinases, their architectural features, how activity is impacted by other proteins, and the implications of these findings for human health and disease.

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Activation loop phosphorylation tunes conformational dynamics underlying Pyk2 tyrosine kinase activation

Cited by 4 publications

References 42 publications

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

Regulation and signaling of the LIM domain kinases

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