scholarly journals Molecular Basis for Ser/Thr Specificity in PKA Signaling

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1548 ◽  
Author(s):  
Matthias J. Knape ◽  
Maximilian Wallbott ◽  
Nicole C. G. Burghardt ◽  
Daniela Bertinetti ◽  
Jan Hornung ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Substrate Specificity ◽  
Metal Ions ◽  
Protein Kinases ◽  
Molecular Basis ◽  
Kinase Inhibitor ◽  
Adenine Nucleotides ◽  
Catalytic Efficiency ◽  
Specific Protein ◽  
Divalent Metal Ions

cAMP-dependent protein kinase (PKA) is the major receptor of the second messenger cAMP and a prototype for Ser/Thr-specific protein kinases. Although PKA strongly prefers serine over threonine substrates, little is known about the molecular basis of this substrate specificity. We employ classical enzyme kinetics and a surface plasmon resonance (SPR)-based method to analyze each step of the kinase reaction. In the absence of divalent metal ions and nucleotides, PKA binds serine (PKS) and threonine (PKT) substrates, derived from the heat-stable protein kinase inhibitor (PKI), with similar affinities. However, in the presence of metal ions and adenine nucleotides, the Michaelis complex for PKT is unstable. PKA phosphorylates PKT with a higher turnover due to a faster dissociation of the product complex. Thus, threonine substrates are not necessarily poor substrates of PKA. Mutation of the DFG+1 phenylalanine to β-branched amino acids increases the catalytic efficiency of PKA for a threonine peptide substrate up to 200-fold. The PKA Cα mutant F187V forms a stable Michaelis complex with PKT and shows no preference for serine versus threonine substrates. Disease-associated mutations of the DFG+1 position in other protein kinases underline the importance of substrate specificity for keeping signaling pathways segregated and precisely regulated.

Divalent metal ions control activity and inhibition of protein kinases

Metallomics ◽  
2017 ◽  
Vol 9 (11) ◽  
pp. 1576-1584 ◽  
Author(s):  
Matthias J. Knape ◽  
Mike Ballez ◽  
Nicole C. Burghardt ◽  
Bastian Zimmermann ◽  
Daniela Bertinetti ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Metal Ions ◽  
Protein Kinases ◽  
Substrate Binding ◽  
Divalent Metal ◽  
Catalytic Cycle ◽  
Divalent Metal Ions ◽  
Control Activity ◽  
Product Release

Metals like Zn2+ and Mn2+ can assist in the catalytic cycle of a protein kinase by facilitating substrate binding and phosphotransfer, however, in contrast to Mg2+ they also hamper product release.

Molecular basis for the substrate specificity of a serine threonine-specific protein kinase

1993 ◽  
Vol 115 (22) ◽  
pp. 9888-9891 ◽  
Author(s):  
Tae Ryong Lee ◽  
Marianne Mendelow ◽  
Jaya Srinivasan ◽  
Young Guen Kwon ◽  
David S. Lawrence
Keyword(s):  
Protein Kinase ◽  
Substrate Specificity ◽  
Molecular Basis ◽  
Specific Protein ◽  
Specific Protein Kinase

scholarly journals Identification of the glycogenic compound 5-iodotubercidin as a general protein kinase inhibitor

1994 ◽  
Vol 299 (1) ◽  
pp. 123-128 ◽  
Author(s):  
D Massillon ◽  
W Stalmans ◽  
G van de Werve ◽  
M Bollen
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Kinase Inhibitor ◽  
Glycogen Synthase ◽  
Biological Effects ◽  
Glycogen Synthesis ◽  
Specific Protein ◽  
Liver Cells ◽  
Protein Kinase Inhibitor ◽  
Casein Kinase 1

Addition of micromolar concentrations of the adenosine derivative 5-iodotubercidin (Itu) initiates glycogen synthesis in isolated hepatocytes by causing inactivation of phosphorylase and activation of glycogen synthase [Flückiger-Isler and Walter (1993) Biochem. J. 292, 85-91]. We report here that Itu also antagonizes the effects of saturating concentrations of glucagon and vasopressin on these enzymes. The Itu-induced activation of glycogen synthase could not be explained by the removal of phosphorylase a (a potent inhibitor of the glycogen-associated synthase phosphatase). When tested on purified enzymes, Itu did not affect the activities of the major Ser/Thr-specific protein phosphatases (PP-1, PP-2A, PP-2B and PP-2C), but it inhibited various Ser/Thr-specific protein kinases as well as the tyrosine kinase activity of the insulin receptor (IC50 between 0.4 and 28 microM at 10-15 microM ATP). Tubercidin, which did not affect glycogen synthase or phosphorylase in liver cells, was 300 times less potent as a protein kinase inhibitor. Kinetic analysis of the inhibition of casein kinase-1 and protein kinase A showed that Itu acts as a competitive inhibitor with respect to ATP, and as a mixed-type inhibitor with respect to the protein substrate. We propose that Itu inactivates phosphorylase and activates glycogen synthase by inhibiting phosphorylase kinase and various glycogen synthase kinases. Consistent with the broad specificity of Itu in vitro, this compound decreased the phosphorylation level of numerous phosphopolypeptides in intact liver cells. Our data suggest that at least some of the biological effects of Itu can be explained by an inhibition of protein kinases.

scholarly journals Betulin, a Newly Characterized Compound in Acacia auriculiformis Bark, Is a Multi-Target Protein Kinase Inhibitor

Molecules ◽  
2021 ◽  
Vol 26 (15) ◽  
pp. 4599
Author(s):  
Augustine Ahmadu ◽  
Claire Delehouzé ◽  
Anas Haruna ◽  
Lukman Mustapha ◽  
Bilqis Lawal ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Kinase Inhibitor ◽  
Glycogen Synthase ◽  
Growth Factor Receptor ◽  
Binding Pocket ◽  
Stem Bark ◽  
Janus Kinase ◽  
Myelogenous Leukemia ◽  
Acacia Auriculiformis

The purpose of this work is to investigate the protein kinase inhibitory activity of constituents from Acacia auriculiformis stem bark. Column chromatography and NMR spectroscopy were used to purify and characterize betulin from an ethyl acetate soluble fraction of acacia bark. Betulin, a known inducer of apoptosis, was screened against a panel of 16 disease-related protein kinases. Betulin was shown to inhibit Abelson murine leukemia viral oncogene homolog 1 (ABL1) kinase, casein kinase 1ε (CK1ε), glycogen synthase kinase 3α/β (GSK-3 α/β), Janus kinase 3 (JAK3), NIMA Related Kinase 6 (NEK6), and vascular endothelial growth factor receptor 2 kinase (VEGFR2) with activities in the micromolar range for each. The effect of betulin on the cell viability of doxorubicin-resistant K562R chronic myelogenous leukemia cells was then verified to investigate its putative use as an anti-cancer compound. Betulin was shown to modulate the mitogen-activated protein (MAP) kinase pathway, with activity similar to that of imatinib mesylate, a known ABL1 kinase inhibitor. The interaction of betulin and ABL1 was studied by molecular docking, revealing an interaction of the inhibitor with the ABL1 ATP binding pocket. Together, these data demonstrate that betulin is a multi-target inhibitor of protein kinases, an activity that can contribute to the anticancer properties of the natural compound and to potential treatments for leukemia.

Molecular Basis for p38 Protein Kinase Inhibitor Specificity

Biochemistry ◽  
1998 ◽  
Vol 37 (47) ◽  
pp. 16573-16581 ◽  
Author(s):  
JeanMarie Lisnock ◽  
Andy Tebben ◽  
Betsy Frantz ◽  
Edward A. O'Neill ◽  
Gist Croft ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Molecular Basis ◽  
Kinase Inhibitor ◽  
Protein Kinase Inhibitor ◽  
Inhibitor Specificity

The Sucrose Non-Fermenting 1-Related Protein Kinase 2 (SnRK2) Genes Are Multifaceted Players in Plant Growth, Development and Response to Environmental Stimuli

2019 ◽  
Vol 61 (2) ◽  
pp. 225-242 ◽  
Author(s):  
Xinguo Mao ◽  
Yuying Li ◽  
Shoaib Ur Rehman ◽  
Lili Miao ◽  
Yanfei Zhang ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Specific Protein ◽  
Regulatory Mechanisms ◽  
Biological Processes ◽  
Related Protein ◽  
Biological Functions ◽  
Reversible Protein Phosphorylation ◽  
Growth Development ◽  
Regulatory Event

Abstract Reversible protein phosphorylation orchestrated by protein kinases and phosphatases is a major regulatory event in plants and animals. The SnRK2 subfamily consists of plant-specific protein kinases in the Ser/Thr protein kinase superfamily. Early observations indicated that SnRK2s are mainly involved in response to abiotic stress. Recent evidence shows that SnRK2s are multifarious players in a variety of biological processes. Here, we summarize the considerable knowledge of SnRK2s, including evolution, classification, biological functions and regulatory mechanisms at the epigenetic, post-transcriptional and post-translation levels.

scholarly journals Potential Role for Protein Kinases in Regulation of Bidirectional Endoplasmic Reticulum-to-Golgi Transport Revealed by Protein Kinase Inhibitor H89

2000 ◽  
Vol 11 (8) ◽  
pp. 2577-2590 ◽  
Author(s):  
Tina H. Lee ◽  
Adam D. Linstedt
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Protein Kinases ◽  
Potential Role ◽  
Kinase Inhibitor ◽  
Transport Processes ◽  
Protein Kinase Inhibitors ◽  
Er Export ◽  
Kinase A

Recent evidence suggests a regulatory connection between cell volume, endoplasmic reticulum (ER) export, and stimulated Golgi-to-ER transport. To investigate the potential role of protein kinases we tested a panel of protein kinase inhibitors for their effect on these steps. One inhibitor, H89, an isoquinolinesulfonamide that is commonly used as a selective protein kinase A inhibitor, blocked both ER export and hypo-osmotic-, brefeldin A-, or nocodazole-induced Golgi-to-ER transport. In contrast, H89 did not block the constitutive ER Golgi-intermediate compartment (ERGIC)-to-ER and Golgi-to-ER traffic that underlies redistribution of ERGIC and Golgi proteins into the ER after ER export arrest. Surprisingly, other protein kinase A inhibitors, KT5720 and H8, as well as a set of protein kinase C inhibitors, had no effect on these transport processes. To test whether H89 might act at the level of either the coatomer protein (COP)I or the COPII coat protein complex we examined the localization of βCOP and Sec13 in H89-treated cells. H89 treatment led to a rapid loss of Sec13-labeled ER export sites but βCOP localization to the Golgi was unaffected. To further investigate the effect of H89 on COPII we developed a COPII recruitment assay with permeabilized cells and found that H89 potently inhibited binding of exogenous Sec13 to ER export sites. This block occurred in the presence of guanosine-5′-O-(3-thio)triphosphate, suggesting that Sec13 recruitment is inhibited at a step independent of the activation of the GTPase Sar1. These results identify a requirement for an H89-sensitive factor(s), potentially a novel protein kinase, in recruitment of COPII to ER export sites, as well as in stimulated but not constitutive Golgi-to-ER transport.

Protein Kinase InhibitorsThe Tyrosine-Specific Protein Kinases

1998 ◽  
Vol 77 (2) ◽  
pp. 81-114 ◽  
Author(s):  
David S. Lawrence ◽  
Jinkui Niu
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Specific Protein

scholarly journals Molecular Basis for p38 Protein Kinase Inhibitor Specificity

Biochemistry ◽  
1999 ◽  
Vol 38 (11) ◽  
pp. 3456-3456
Author(s):  
JeanMarie Lisnock ◽  
Andy Tebben ◽  
Betsy Frantz ◽  
Edward A. O'Neill ◽  
Gist Croft ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Molecular Basis ◽  
Kinase Inhibitor ◽  
Protein Kinase Inhibitor ◽  
Inhibitor Specificity
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