The Crystal Structure of Cancer Osaka Thyroid Kinase Reveals an Unexpected Kinase Domain Fold

Gutmann, Sascha; Hinniger, Alexandra; Fendrich, Gabriele; Drückes, Peter; Antz, Sylvie; Mattes, Henri; Möbitz, Henrik; Ofner, Silvio; Schmiedeberg, Niko; Stojanović, Aleksandar; Rieffel, S.; Strauss, André; Troxler, Thomas; Glatthar, Ralf; Sparrer, Helmut

doi:10.1074/jbc.m115.648097

Cited by 26 publications

(27 citation statements)

References 39 publications

(44 reference statements)

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“…Phosphorylation of MKK1 and MKK6 by recombinant TPL-2 protein was determined using 50 nM StrepII-TPL-2 30-404 [24,25] with 400 nM His6-MKK1 D208A and MBP-MKK6 K82A substrates, respectively. Assays were performed in kinase buffer [50 mM HEPES (pH 7.4), 1 mM TCEP, 0.01% BSA, 0.01% Brij35, 5 mM MnCl 2 and 10% DMSO], supplemented with 300 µM ATP ± 10 µM C34 TPL-2 inhibitor [26] for 30 min at 30°C.…”

Section: Methodsmentioning

confidence: 99%

TLR and TNF-R1 activation of the MKK3/MKK6–p38α axis in macrophages is mediated by TPL-2 kinase

Pattison

Mitchell

Flynn

et al. 2016

Biochemical Journal

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Previous studies suggested that Toll-like receptor (TLR) stimulation of the p38α MAP kinase (MAPK) is mediated by transforming growth factor-β-activated kinase 1 (TAK1) activation of MAPK kinases, MKK3, MKK4 and MKK6. We used quantitative mass spectrometry to monitor tumour progression locus 2 (TPL-2)-dependent protein phosphorylation following TLR4 stimulation with lipopolysaccharide, comparing macrophages from wild-type mice and Map3k8D270A/D270A mice expressing catalytically inactive TPL-2 (MAP3K8). In addition to the established TPL-2 substrates MKK1/2, TPL-2 kinase activity was required to phosphorylate the activation loops of MKK3/6, but not of MKK4. MKK3/6 activation required IκB kinase (IKK) phosphorylation of the TPL-2 binding partner nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB1) p105, similar to MKK1/2 activation. Tumour necrosis factor (TNF) stimulation of MKK3/6 phosphorylation was similarly dependent on TPL-2 catalytic activity and IKK phosphorylation of NF-κB1 p105. Owing to redundancy of MKK3/6 with MKK4, Map3k8D270A mutation only fractionally decreased lipopolysaccharide activation of p38α. TNF activation of p38α, which is mediated predominantly via MKK3/6, was substantially reduced. TPL-2 catalytic activity was also required for MKK3/6 and p38α activation following macrophage stimulation with Mycobacterium tuberculosis and Listeria monocytogenes. Our experiments demonstrate that the IKK/NF-κB1 p105/TPL-2 signalling pathway, downstream of TAK1, regulates MKK3/6 and p38α activation in macrophages in inflammation.

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Section: Methodsmentioning

confidence: 99%

TLR and TNF-R1 activation of the MKK3/MKK6–p38α axis in macrophages is mediated by TPL-2 kinase

Pattison

Mitchell

Flynn

et al. 2016

Biochemical Journal

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“…TPL-2 30–404 protein was produced following a published methodology [ 9 ]. Sf9 cells were co-transfected with pFastbac virus DNA for baculovirus generation.…”

Section: Methodsmentioning

confidence: 99%

“…Protein expression was carried out in 5-l cell cultures of Sf9 cells with plaque-purified viruses. Compound 1 (10 µM; [ 9 ]) was added to the cell cultures at 24 h to improve recombinant TPL-2 30–404 protein yields. Cells were harvested 72 h after infection, pellets were snap-frozen and stored at −80°C.…”

Section: Methodsmentioning

confidence: 99%

“…TPL-2 is a MKK 1/2 kinase, which mediates activation of ERK1/2 MAP kinases by TLR4 (and all other TLRs), as well as the receptors for TNF and IL-1β, in macrophages [ 5 ]. Several pharmaceutical companies have carried out screens using the isolated TPL-2 kinase domain and MKK1 substrate to identify inhibitors that block TPL-2 activation of ERK1/2 in cultured macrophages [ 8 , 9 ]. However, none of these have progressed to clinical trial, likely reflecting the challenge of producing ATP-competitive inhibitors with the appropriate physicochemical characteristics required for drug development.…”

Section: Introductionmentioning

confidence: 99%

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Assaying kinase activity of the TPL-2/NF-κB1 p105/ABIN-2 complex using an optimal peptide substrate

Kümper

Gantke

Chen

et al. 2018

Biochemical Journal

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The MKK1/2 kinase tumour progression locus 2 (TPL-2) is critical for the production of tumour necrosis factor alpha (TNFα) in innate immune responses and a potential anti-inflammatory drug target. Several earlier pharmaceutical company screens with the isolated TPL-2 kinase domain have identified small-molecule inhibitors that specifically block TPL-2 signalling in cells, but none of these have progressed to clinical development. We have previously shown that TPL-2 catalytic activity regulates TNF production by macrophages while associated with NF-κB1 p105 and ABIN-2, independently of MKK1/2 phosphorylation via an unknown downstream substrate. In the present study, we used a positional scanning peptide library to determine the optimal substrate specificity of a complex of TPL-2, NF-κB1 p105 and ABIN-2. Using an optimal peptide substrate based on this screen and a high-throughput mass spectrometry assay to monitor kinase activity, we found that the TPL-2 complex has significantly altered sensitivities versus existing ATP-competitive TPL-2 inhibitors than the isolated TPL-2 kinase domain. These results imply that screens with the more physiologically relevant TPL-2/NF-κB1 p105/ABIN-2 complex have the potential to deliver novel TPL-2 chemical series; both ATP-competitive and allosteric inhibitors could emerge with significantly improved prospects for development as anti-inflammatory drugs.

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“…19 There are so many inhibitors have been identified to reduce the harmful effect of the COT kinase such as (E)-3-(2-amino-5 -( n a p h t h a l e n -2 -y l ) p y r i d i n -3 -y l ) a c r y l i c a c i d , 5-(5-(1H-indol-3-yl)-1H-pyrrolo [2,3-b]pyridin-3-yl)-1,3,4-oxadiazol-2-amine,5-(2-amino-5-(quinolin-3-yl)pyridin-3-yl)-1,3,4-oxadiazole-2(3H)-thione, 20 4-alkylamino-(1,7) naphthyridine-3-carbonitriles, 21 Indazoles, 22 1,7-naphthy ridine-3-carbonitriles, 23 thieno (2, 3-c) pyridine, 24,25 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles. 26 So there is a need to find out the small molecule cancer drugs.…”

mentioning

confidence: 99%

A Combination of Pharmacophore Modeling, Molecular Docking and Virtual Screening Study Reveals 3,5,7-Trihydroxy-2-(3,4,5-trihydroxyphenyl)- 4H-Chromen-4-One as a Potential Anti-Cancer Agent of COT Kinase

Hussain¹,

Verma²

2018

IJPER

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Background: COT (Tpl2/MAP3K8) is a Serine/ Threonine protein kinase which plays a crucial role in the production of TNF-alpha through the phosphorylation of MEK, ERK pathway and the production of other pro-inflammatory cytokines. Its inhibition has been shown as important to reduce inflammatory diseases and cancer. Material and Methods: Combined Ligand-based and Structure-based pharmacophore model was developed for finding out the potential anticancer agents. These combined pharmacophore model was used as 3D-query for searching the matching pharmacophore features against chemical structure databases such as DrugBank, MDPI, ZINC, Maybridge HitFinder. Docking was performed using Schrodinger software as well as selected Hits were filtered by ADMET properties. Results: Among all the selected Hits Compound 3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)-4H-chromen-4-one was found to be more potent according to the docking score. Conclusion: A step by step computational pipeline was used to find out the potential anticancer agents. This study suggests that these Hits could be used as anticancer agents against death leading diseases.

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The Crystal Structure of Cancer Osaka Thyroid Kinase Reveals an Unexpected Kinase Domain Fold

Cited by 26 publications

References 39 publications

TLR and TNF-R1 activation of the MKK3/MKK6–p38α axis in macrophages is mediated by TPL-2 kinase

TLR and TNF-R1 activation of the MKK3/MKK6–p38α axis in macrophages is mediated by TPL-2 kinase

Assaying kinase activity of the TPL-2/NF-κB1 p105/ABIN-2 complex using an optimal peptide substrate

A Combination of Pharmacophore Modeling, Molecular Docking and Virtual Screening Study Reveals 3,5,7-Trihydroxy-2-(3,4,5-trihydroxyphenyl)- 4H-Chromen-4-One as a Potential Anti-Cancer Agent of COT Kinase

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