scholarly journals Sequence of pp42/MAP kinase, a serine/threonine kinase regulated by tyrosine phosphorylation

1991 ◽  
Vol 19 (13) ◽  
pp. 3743-3743 ◽  
Author(s):  
Jeng-Horng Her ◽  
Jie Wu ◽  
Thomas B. Rall ◽  
Thomas W. Sturgill ◽  
Michael J. Weber
Keyword(s):  
Map Kinase ◽  
Tyrosine Phosphorylation ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Kinase A

Development of a Transcreener™ Kinase Assay for Protein Kinase A and Demonstration of Concordance of Data with a Filter-Binding Assay Format

2007 ◽  
Vol 12 (4) ◽  
pp. 578-584 ◽  
Author(s):  
Karen L. Huss ◽  
Pauline E. Blonigen ◽  
Robert M. Campbell
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Fluorescence Polarization ◽  
Binding Assay ◽  
Kinase Assay ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Alexa Fluor ◽  
Small Molecule Library ◽  
Kinase A

A Transcreener™ kinase fluorescence polarization (FP) assay has been developed for the serine/threonine kinase protein kinase A (PKA). The PKA Transcreener™ kinase assay is an homogenous, competitive antibody-based FP assay that uses Far Red Alexa Fluor 633-labeled adenosine 5′ disphosphate (ADP) tracer and mouse monoclonal anti-ADP antibody. The Transcreener™ PKA assay was validated with both known PKA inhibitors and library compounds. The Transcreener™ PKA assay is resistant to low-wavelength (or common) fluorescent interference from small-molecule library compounds and generates IC50 results comparable with current radioactive filter-binding assay. ( Journal of Biomolecular Screening 2007:578-584)

scholarly journals Fyn Regulates Binding Partners of Cyclic-AMP Dependent Protein Kinase A

Proteomes ◽  
2018 ◽  
Vol 6 (4) ◽  
pp. 37
Author(s):  
Anna Schmoker ◽  
Samuel Barritt ◽  
Marion Weir ◽  
Jacqueline Mann ◽  
Tyler Hogan ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Quantitative Mass Spectrometry ◽  
Serine Threonine Kinase ◽  
Dependent Protein Kinase ◽  
Threonine Kinase ◽  
Cellular Processes ◽  
Binding Partners ◽  
Dependent Protein ◽  
Kinase A

The cAMP-dependent protein kinase A (PKA) is a serine/threonine kinase involved in many fundamental cellular processes, including migration and proliferation. Recently, we found that the Src family kinase Fyn phosphorylates the catalytic subunit of PKA (PKA-C) at Y69, thereby increasing PKA kinase activity. We also showed that Fyn induced the phosphorylation of cellular proteins within the PKA preferred target motif. This led to the hypothesis that Fyn could affect proteins in complex with PKA. To test this, we employed a quantitative mass spectrometry approach to identify Fyn-dependent binding partners in complex with PKA-C. We found Fyn enhanced the binding of PKA-C to several cytoskeletal regulators that localize to the centrosome and Golgi apparatus. Three of these Fyn-induced PKA interactors, AKAP9, PDE4DIP, and CDK5RAP2, were validated biochemically and were shown to exist in complex with Fyn and PKA in a glioblastoma cell line. Intriguingly, the complexes formed between PKA-C and these known AKAPs were dependent upon Fyn catalytic activity and expression levels. In addition, we identified Fyn-regulated phosphorylation sites on proteins in complex with PKA-C. We also identified and biochemically validated a novel PKA-C interactor, LARP4, which complexed with PKA in the absence of Fyn. These results demonstrate the ability of Fyn to influence the docking of PKA to specific cellular scaffolds and suggest that Fyn may affect the downstream substrates targeted by PKA.

Interleukin-4-induced transcriptional activation by Stat6 involves multiple serine/threonine kinase pathways and serine phosphorylation of Stat6

Blood ◽  
2000 ◽  
Vol 95 (2) ◽  
pp. 494-502 ◽  
Author(s):  
Marko Pesu ◽  
Kati Takaluoma ◽  
Saara Aittomäki ◽  
Anssi Lagerstedt ◽  
Kalle Saksela ◽  
...  
Keyword(s):  
Dna Binding ◽  
Tyrosine Phosphorylation ◽  
Transcriptional Activation ◽  
Kinase Inhibitor ◽  
Serine Phosphorylation ◽  
Interleukin 4 ◽  
Specific Gene ◽  
Serine Threonine Kinase ◽  
Transcriptional Responses ◽  
Threonine Kinase

Stat6 transcription factor is a critical mediator of IL-4-specific gene responses. Tyrosine phosphorylation is required for nuclear localization and DNA binding of Stat6. The authors investigated whether Stat6-dependent transcriptional responses are regulated through IL-4-induced serine/threonine phosphorylation. In Ramos B cells, the serine/threonine kinase inhibitor H7 inhibited IL-4-induced expression of CD23. Treatment with H7 did not affect IL-4R-mediated immediate signaling events such as tyrosine phosphorylation of Jak1, Jak3, insulin receptor substrate (IRS)-1 and IRS-2, or tyrosine phosphorylation and DNA binding of Stat6. To analyze whether the H7-sensitive pathway was regulating Stat6-activated transcription, we used reporter constructs containing different IL-4 responsive elements. H7 abrogated Stat6-, as well as Stat5-, mediated reporter gene activation and partially reduced C/EBP-dependent reporter activity. By contrast, IL-4-induced transcription was not affected by wortmannin, an inhibitor of the phosphatidyl-inositol 3′-kinase pathway. Phospho-amino acid analysis and tryptic phosphopeptide maps revealed that IL-4 induced phosphorylation of Stat6 on serine and tyrosine residues in Ramos cells and in 32D cells lacking endogenous IRS proteins. However, H7 treatment did not inhibit the phosphorylation of Stat6. Instead, H7 inhibited the IL-4-induced phosphorylation of RNA polymerase II. These results indicate that Stat6-induced transcription is dependent on phosphorylation events mediated by H7-sensitive kinase(s) but that it also involves serine phosphorylation of Stat6 by an H7-insensitive kinase independent of the IRS pathway.

scholarly journals Inhibition of Bcr serine kinase by tyrosine phosphorylation.

1996 ◽  
Vol 16 (3) ◽  
pp. 998-1005 ◽  
Author(s):  
J Liu ◽  
Y Wu ◽  
G Z Ma ◽  
D Lu ◽  
L Haataja ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Tyrosine Phosphorylation ◽  
Kinase Activity ◽  
Philadelphia Chromosome ◽  
Wild Type ◽  
Serine Kinase ◽  
Serine Threonine Kinase ◽  
Threonine Kinase

The first exon of the BCR gene encodes a new serine/threonine protein kinase. Abnormal fusion of the BCR and ABL genes, resulting from the formation of the Philadelphia chromosome (Ph), is the hallmark of Ph-positive leukemia. We have previously demonstrated that the Bcr protein is tyrosine phosphorylated within first-exon sequences by the Bcr-Abl oncoprotein. Here we report that in addition to tyrose 177 (Y-177), Y-360 and Y283 are phosphorylated in Bcr-Abl proteins in vitro. Moreover, Bcr tyrosine 360 is phosphorylated in vivo within both Bcr-Abl and Bcr. Bcr mutant Y177F had a greatly reduced ability to transphosphorylate casein and histone H1, whereas Bcr mutants Y177F and Y283F had wild-type activities. In contrast, the Y360F mutation had little effect on Bcr's autophosphorylation activity. Tyrosine-phosphorylated Bcr, phosphorylated in vitro by Bcr-Abl, was greatly inhibited in its serine/threonine kinase activity, impairing both auto- and transkinase activities of Bcr. Similarly, the isolation of Bcr from cells expressing Bcr-Abl under conditions that preserve phosphotyrosine residues also reduced Bcr's kinase activity. These results indicate that tyrosine 360 of Bcr is critical for the transphosphorylation activity of Bcr and that in Ph-positive leukemia, Bcr serine/threonine kinase activity is seriously impaired.

scholarly journals Rho-associated kinase, a novel serine/threonine kinase, as a putative target for small GTP binding protein Rho.

1996 ◽  
Vol 15 (9) ◽  
pp. 2208-2216 ◽  
Author(s):  
T. Matsui ◽  
M. Amano ◽  
T. Yamamoto ◽  
K. Chihara ◽  
M. Nakafuku ◽  
...  
Keyword(s):  
Binding Protein ◽  
Serine Threonine Kinase ◽  
Gtp Binding Protein ◽  
Threonine Kinase ◽  
Gtp Binding ◽  
Putative Target ◽  
Kinase A

scholarly journals Tyrosine Phosphorylation of A17 during Vaccinia Virus Infection: Involvement of the H1 Phosphatase and the F10 Kinase

1999 ◽  
Vol 73 (9) ◽  
pp. 7287-7296 ◽  
Author(s):  
M. Derrien ◽  
A. Punjabi ◽  
M. Khanna ◽  
O. Grubisha ◽  
P. Traktman
Keyword(s):  
Virus Infection ◽  
Vaccinia Virus ◽  
Tyrosine Phosphorylation ◽  
Temperature Sensitive ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Tyrosine Residues ◽  
Vaccinia Virus Infection ◽  
Dual Specificity ◽  
Phosphotyrosine Signaling

ABSTRACT Vaccinia virus encodes two protein kinases (B1 and F10) and a dual-specificity phosphatase (VH1), suggesting that phosphorylation and dephosphorylation of substrates on serine/threonine and tyrosine residues are important in regulating diverse aspects of the viral life cycle. Using a recombinant in which expression of the H1 phosphatase can be regulated experimentally (vindH1), we have previously demonstrated that repression of H1 leads to the maturation of noninfectious virions that contain several hyperphosphorylated substrates (K. Liu et al., J. Virol. 69:7823–7834). In this report, we demonstrate that among these is a 25-kDa protein that is phosphorylated on tyrosine residues in H1-deficient virions and can be dephosphorylated by recombinant H1. We demonstrate that the 25-kDa phosphoprotein represents the product of the A17 gene and that A17 is phosphorylated on serine, threonine, and tyrosine residues during infection. Detection of phosphotyrosine within A17 is abrogated when Tyr203 (but not Tyr3, Tyr6, or Tyr7) is mutated to phenylalanine, suggesting strongly that this amino acid is the site of tyrosine phosphorylation. Phosphorylation of A17 fails to occur during nonpermissive infections performed with temperature-sensitive mutants defective in the F10 kinase. Our data suggest that this enzyme, which was initially characterized as a serine/threonine kinase, might in fact have dual specificity. This hypothesis is strengthened by the observation thatEscherichia coli induced to express F10 contain multiple proteins which are recognized by antiphosphotyrosine antiserum. This study presents the first evidence for phosphotyrosine signaling during vaccinia virus infection and implicates the F10 kinase and the H1 phosphatase as the dual-specificity enzymes that direct this cycle of reversible phosphorylation.

scholarly journals cRel-TD kinase: a serine/threonine kinase binding in vivo and in vitro c-Rel and phosphorylating its transactivation domain

Oncogene ◽  
2000 ◽  
Vol 19 (18) ◽  
pp. 2224-2232 ◽  
Author(s):  
Caterina Fognani ◽  
Rossana Rondi ◽  
Alfredo Romano ◽  
Francesco Blasi
Keyword(s):  
Transactivation Domain ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Kinase A
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