scholarly journals PKA-phosphorylation of PDE4D3 facilitates recruitment of the mAKAP signalling complex

2004 ◽  
Vol 381 (3) ◽  
pp. 587-592 ◽  
Author(s):  
Jennifer J. CARLISLE MICHEL ◽  
Kimberly L. DODGE ◽  
Wei WONG ◽  
Nicole C. MAYER ◽  
Lorene K. LANGEBERG ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Negative Feedback ◽  
Feedback Loop ◽  
Negative Feedback Loop ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Anchoring Protein ◽  
Dependent Protein

mAKAP (muscle-selective A-kinase-anchoring protein) co-ordinates a cAMP-sensitive negative-feedback loop comprising PKA (cAMP-dependent protein kinase) and the cAMP-selective PDE4D3 (phosphodiesterase 4D3). In vitro and cellular experiments demonstrate that PKA-phosphorylation of PDE4D3 on Ser-13 increases the affinity of PDE4D3 for mAKAP. Our data suggest that activation of mAKAP-anchored PKA enhances the recruitment of PDE4D3, allowing for quicker signal termination.

Lipopolysaccharide‐responsive beige‐like anchor acts as a cAMP‐dependent protein kinase anchoring protein in B cells

2020 ◽  
Vol 92 (3) ◽  
Author(s):  
Nidia Carolina Moreno‐Corona ◽  
Orestes Lopez‐Ortega ◽  
Jose Mizael Flores Hermenegildo ◽  
Laura Berron‐Ruiz ◽  
Juan Carlos Rodriguez‐Alba ◽  
...  
Keyword(s):  
B Cells ◽  
Protein Kinase ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Anchoring Protein ◽  
Dependent Protein

scholarly journals A CaMKII/PDE4D negative feedback regulates cAMP signaling

2015 ◽  
Vol 112 (7) ◽  
pp. 2023-2028 ◽  
Author(s):  
Delphine Mika ◽  
Wito Richter ◽  
Marco Conti
Keyword(s):  
Protein Kinase ◽  
Negative Feedback ◽  
Feedback Loop ◽  
Dependent Protein Kinase ◽  
Protein Targets ◽  
Cardiac Pathophysiology ◽  
Functional Consequences ◽  
Dependent Protein ◽  
Cardiac Excitation ◽  
Camp Levels

cAMP production and protein kinase A (PKA) are the most widely studied steps in β-adrenergic receptor (βAR) signaling in the heart; however, the multifunctional Ca2+/calmodulin-dependent protein kinase II (CaMKII) is also activated in response to βAR stimulation and is involved in the regulation of cardiac excitation-contraction coupling. Its activity and expression are increased during cardiac hypertrophy, in heart failure, and under conditions that promote arrhythmias both in animal models and in the human heart, underscoring the clinical relevance of CaMKII in cardiac pathophysiology. Both CaMKII and PKA phosphorylate a number of protein targets critical for Ca2+ handling and contraction with similar, but not always identical, functional consequences. How these two pathways communicate with each other remains incompletely understood, however. To maintain homeostasis, cyclic nucleotide levels are regulated by phosphodiesterases (PDEs), with PDE4s predominantly responsible for cAMP degradation in the rodent heart. Here we have reassessed the interaction between cAMP/PKA and Ca2+/CaMKII signaling. We demonstrate that CaMKII activity constrains basal and βAR-activated cAMP levels. Moreover, we show that these effects are mediated, at least in part, by CaMKII regulation of PDE4D. This regulation establishes a negative feedback loop necessary to maintain cAMP/CaMKII homeostasis, revealing a previously unidentified function for PDE4D as a critical integrator of cAMP/PKA and Ca2+/CaMKII signaling.

scholarly journals Regulation of Microtubule Dynamics by Extracellular Signals: cAMP-dependent Protein Kinase Switches Off the Activity of Oncoprotein 18 in Intact Cells

1998 ◽  
Vol 140 (1) ◽  
pp. 131-141 ◽  
Author(s):  
Helena Melander Gradin ◽  
Niklas Larsson ◽  
Ulrica Marklund ◽  
Martin Gullberg
Keyword(s):  
Protein Kinase ◽  
Genetic System ◽  
Cell Surface Receptor ◽  
In Vitro Assays ◽  
Dependent Protein Kinase ◽  
Intact Cells ◽  
Camp Dependent Protein Kinase ◽  
Oncoprotein 18 ◽  
Dependent Protein

Oncoprotein 18 (Op18, also termed p19, 19K, metablastin, stathmin, and prosolin) is a recently identified regulator of microtubule (MT) dynamics. Op18 is a target for both cell cycle and cell surface receptor-coupled kinase systems, and phosphorylation of Op18 on specific combinations of sites has been shown to switch off its MT-destabilizing activity. Here we show that induced expression of the catalytic subunit of cAMP-dependent protein kinase (PKA) results in a dramatic increase in cellular MT polymer content concomitant with phosphorylation and partial degradation of Op18. That PKA may regulate the MT system by downregulation of Op18 activity was evaluated by a genetic system allowing conditional co-expression of PKA and a series of kinase target site–deficient mutants of Op18. The results show that phosphorylation of Op18 on two specific sites, Ser-16 and Ser-63, is necessary and sufficient for PKA to switch off Op18 activity in intact cells. The regulatory importance of dual phosphorylation on Ser-16 and Ser-63 of Op18 was reproduced by in vitro assays. These results suggest a simple model where PKA phosphorylation downregulates the MT-destabilizing activity of Op18, which in turn promotes increased tubulin polymerization. Hence, the present study shows that Op18 has the potential to regulate the MT system in response to external signals such as cAMP-linked agonists.

scholarly journals Adrenocorticotrophic hormone stimulates phosphotyrosine phosphatase SHP2 in bovine adrenocortical cells: phosphorylation and activation by cAMP-dependent protein kinase

2000 ◽  
Vol 352 (2) ◽  
pp. 483-490 ◽  
Author(s):  
Stéphane ROCCHI ◽  
Isabelle GAILLARD ◽  
Emmanuel VAN OBBERGHEN ◽  
Edmond M. CHAMBAZ ◽  
Isabelle VILGRAIN
Keyword(s):  
Protein Kinase ◽  
Kinase Assay ◽  
Dependent Manner ◽  
Adrenocorticotrophic Hormone ◽  
Adrenocortical Cells ◽  
Dependent Protein Kinase ◽  
Phosphotyrosine Phosphatase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

During activation of adrenocortical cells by adrenocorticotrophic hormone (ACTH), tyrosine dephosphorylation of paxillin is stimulated and this correlates with protrusion of filopodial structures and a decreased number of focal adhesions. These effects are inhibited by Na3VO4, a phosphotyrosine phosphatase inhibitor [Vilgrain, Chinn, Gaillard, Chambaz and Feige (1998) Biochem. J. 332, 533–540]. However, the tyrosine phosphatases involved in these processes remain to be identified. In this study, we provide evidence that the Src homology domain (SH)2-containing phosphotyrosine phosphatase (SHP)2, but not SHP1, is expressed in adrenocortical cells and is phosphorylated upon ACTH challenge. ACTH (10-8M) treatment of 32P-labelled adrenocortical cells resulted in an increase in phosphorylated SHP2. By probing SHP2-containing immunoprecipitates with an antibody to phosphoserine we found that SHP2 was phosphorylated on serine in ACTH-treated cells in a dose- and time-dependent manner. Furthermore, using an in vitro kinase assay, we showed that SHP2 was a target for cAMP-dependent protein kinase (PKA). Serine was identified as the only target amino acid phosphorylated in SHP2. Phosphorylation of SHP2 by PKA resulted in a dramatic stimulation of phosphatase activity measured either with insulin receptor substrate-1 or with the synthetic peptide [32P]poly(Glu/Tyr) as substrate. In an in-gel assay of SHP2-containing immunoprecipitates, phosphorylated in vitro by PKA or isolated from adrenocortical cells treated with 10nM ACTH, a pronounced activation of SHP2 activity was shown. These observations clearly support the idea that a PKA-mediated signal transduction pathway contributes to SHP2 regulation in adrenocortical cells and point to SHP2 as a possible mediator of the effects of ACTH.

scholarly journals cAMP-dependent protein kinase (PKA) complexes probed by complementary differential scanning fluorimetry and ion mobility–mass spectrometry

2016 ◽  
Vol 473 (19) ◽  
pp. 3159-3175 ◽  
Author(s):  
Dominic P. Byrne ◽  
Matthias Vonderach ◽  
Samantha Ferries ◽  
Philip J. Brownridge ◽  
Claire E. Eyers ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Kinase ◽  
Ion Mobility ◽  
Catalytic Domain ◽  
Ion Mobility Mass Spectrometry ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Differential Scanning Fluorimetry ◽  
Dependent Protein

cAMP-dependent protein kinase (PKA) is an archetypal biological signaling module and a model for understanding the regulation of protein kinases. In the present study, we combine biochemistry with differential scanning fluorimetry (DSF) and ion mobility–mass spectrometry (IM–MS) to evaluate effects of phosphorylation and structure on the ligand binding, dynamics and stability of components of heteromeric PKA protein complexes in vitro. We uncover dynamic, conformationally distinct populations of the PKA catalytic subunit with distinct structural stability and susceptibility to the physiological protein inhibitor PKI. Native MS of reconstituted PKA R2C2 holoenzymes reveals variable subunit stoichiometry and holoenzyme ablation by PKI binding. Finally, we find that although a ‘kinase-dead’ PKA catalytic domain cannot bind to ATP in solution, it interacts with several prominent chemical kinase inhibitors. These data demonstrate the combined power of IM–MS and DSF to probe PKA dynamics and regulation, techniques that can be employed to evaluate other protein-ligand complexes, with broad implications for cellular signaling.

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