A Role for the Tyrosine Kinase Pyk2 in Depolarization-induced Contraction of Vascular Smooth Muscle

Mills, Ryan D.; Mita, Mitsuo; Nakagawa, Junichi; Miyake, Shoji; Sutherland, Cindy; Walsh, Michael P.

doi:10.1074/jbc.m114.633107

Cited by 37 publications

(57 citation statements)

References 29 publications

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“…Previous studies have shown that calcium can activate PYK2 by causing autophosphorylation at Tyr-402, a site that is required for kinase activity (33). Active PYK2 in vascular smooth muscle has been shown to mediate Ca 2+ /calmodulin-dependent PYK2 homodimer formation and transphosphorylation (35) and inhibit MYPT1 through a RhoA/ROCK pathway (36). Moreover, PYK2 inhibition has been shown to prevent stretch-induced activation of Akt and ERK1/2 (37).…”

Section: Discussionmentioning

confidence: 99%

CF airway smooth muscle transcriptome reveals a role for PYK2

Cook¹,

Adam

Zarei

et al. 2017

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Section: Discussionmentioning

confidence: 99%

CF airway smooth muscle transcriptome reveals a role for PYK2

Cook¹,

Adam

Zarei

et al. 2017

JCI Insight

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“…-induced rat caudal arterial smooth muscle contraction in a concentration-dependent manner, and pretreatment of the tissue with salicylate eliminated the tonic component of depolarization-induced contraction without affecting the phasic contractile response (Fig. 4a) (Mills et al 2015). Likewise, salicylate abolished the sustained elevation of LC 20 phosphorylation without affecting the phasic K ?…”

Section: Identification Of the Tyrosine Kinase As Pyk2mentioning

confidence: 96%

A role for the Ca2+-dependent tyrosine kinase Pyk2 in tonic depolarization-induced vascular smooth muscle contraction

Mills

Mita

Walsh

2015

J Muscle Res Cell Motil

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Depolarization of the plasma membrane is a key mechanism of activation of contraction of vascular smooth muscle. This is commonly achieved in isolated, de-endothelialized vascular smooth muscle strips by increasing extracellular [K(+)] (replacing Na(+) by K(+)) and leads to a rapid phasic contraction followed by a sustained tonic contraction. The initial phasic contractile response is due to opening of voltage-gated Ca(2+) channels and entry of extracellular Ca(2+), which binds to calmodulin, leading to activation of myosin light chain kinase, phosphorylation of the regulatory light chains of myosin II at Ser19 and cross-bridge cycling. The subsequent tonic contractile response involves, in addition to myosin light chain kinase activation, Ca(2+)-induced Ca(2+) sensitization whereby Ca(2+) entry activates the RhoA/Rho-associated kinase pathway leading to phosphorylation of MYPT1 (the myosin targeting subunit of myosin light chain phosphatase) and inhibition of the phosphatase. Investigations into the mechanism of activation of RhoA by Ca(2+) have implicated a genistein-sensitive tyrosine kinase, and recent evidence indicates this to be the Ca(2+)-dependent tyrosine kinase, Pyk2.

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“…Protein was extracted from each lyophilized tissue in 1 ml of SDS-gel sample buffer with constant shaking for 16 hours at 4°C. Tissue proteins were resolved by Phos-Tag SDS-PAGE and western blotting for the measurement of LC 20 , Par-4 and CPI-17 phosphorylation (Takeya et al, 2008;Moffat et al, 2011;Ihara et al, 2015), and by SDS-PAGE and western blotting with phosphospecific antibodies for MYPT1 phosphorylation (Grassie et al, 2012;Mills et al, 2015). For CPI-17, 12.5% acrylamide and 30 mM Phos-Tag reagent were used, and proteins were transferred to polyvinylidene difluoride (Roche Life Science, Quebec, Canada) overnight at 25 V and 4°C in 10 mM CAPS, pH 11, 10% methanol.…”

Section: Methodsmentioning

confidence: 99%

“…RhoA/ ROCK activation through Ga q/11 can result in contractile responses to PE; however, the functional linkages are incompletely understood (Puetz et al, 2009;Wettschureck and Offermanns, 2005). Moreover, KCl-induced membrane depolarization also elicits sustained contraction of VSM via activation of the RhoA/ROCK pathway (Mita et al, 2002(Mita et al, , 2013Sakamoto et al, 2003;Sakurada et al, 2003;Urban et al, 2003;Mills et al, 2015). The effects of inhibition of ZIPK during agonist-dependent contractions of caudal arterial smooth muscle suggest that the kinase lies downstream from GPCRs that signal through both Ga 12/13 and Ga q/11 (Fig.…”

mentioning

confidence: 99%

A Small Molecule Pyrazolo[3,4-d]Pyrimidinone Inhibitor of Zipper-Interacting Protein Kinase Suppresses Calcium Sensitization of Vascular Smooth Muscle

et al. 2015

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A novel inhibitor of zipper-interacting protein kinase (ZIPK) was used to examine the involvement of ZIPK in the regulation of smooth muscle contraction. Pretreatment of de-endothelialized rat caudal arterial smooth muscle strips with the pyrazolo [3,4-propanamide (HS38) decreased the velocity of contraction (time to reach half-maximal force) induced by the phosphatase inhibitor calyculin A in the presence of Ca 21 without affecting maximal force development. This effect was reversed following washout of HS38 and correlated with a reduction in the rate of phosphorylation of myosin 20-kDa regulatory light chains (LC 20 ) but not of protein kinase C-potentiated inhibitory protein for myosin phosphatase of 17 kDa (CPI-17), prostate apoptosis response-4, or myosin phosphatase-targeting subunit 1 (MYPT1), all of which have been implicated in the regulation of vascular contractility. A structural analog of HS38, with inhibitory activity toward proviral integrations of Moloney (PIM) virus 3 kinase but not ZIPK, had no effect on calyculin A-induced contraction or protein phosphorylations. We conclude that a pool of constitutively active ZIPK is involved in regulation of vascular smooth muscle contraction through direct phosphorylation of LC 20 upon inhibition of myosin light chain phosphatase activity. HS38 also significantly attenuated both phasic and tonic contractile responses elicited by phenylephrine, angiotensin II, endothelin-1, U46619, and K 1 -induced membrane depolarization in the presence of Ca 21 , which correlated with inhibition of phosphorylation of LC 20 , MYPT1, and CPI-17. These effects of HS38 suggest that ZIPK also lies downstream from G protein-coupled receptors that signal through both Ga 12/13 and Ga q/11 .

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A Role for the Tyrosine Kinase Pyk2 in Depolarization-induced Contraction of Vascular Smooth Muscle

Cited by 37 publications

References 29 publications

CF airway smooth muscle transcriptome reveals a role for PYK2

CF airway smooth muscle transcriptome reveals a role for PYK2

A role for the Ca2+-dependent tyrosine kinase Pyk2 in tonic depolarization-induced vascular smooth muscle contraction

A Small Molecule Pyrazolo[3,4-d]Pyrimidinone Inhibitor of Zipper-Interacting Protein Kinase Suppresses Calcium Sensitization of Vascular Smooth Muscle

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