Takinib Inhibits Inflammation in Human Rheumatoid Arthritis Synovial Fibroblasts by Targeting the Janus Kinase-Signal Transducer and Activator of Transcription 3 (JAK/STAT3) Pathway
Abstract
:1. Introduction
2. Results
2.1. Inhibition of TAK1 Reduces Proinflammatory Mediators Secreted by RASFs and THP-1 Monocyte-Derived Macrophages
2.2. Takinib Inhibits STAT3 and JNK Phosphorylation in IL-1β, but Not IL-6 Stimulated Human RASFs
2.3. Takinib Inhibits STAT3 Nuclear Translocation and DNA-Binding Activity in Human RASFs
2.4. Takinib Phosphorylates TAK1 and Fails to Inhibit NF-κB or MAPK Signaling in LPS-Stimulated THP-1 Macrophages
2.5. Docking Simulations Using Takinib and STAT3 Demonstrate a Potential Site of Interaction
3. Discussion
4. Materials and Methods
4.1. Antibodies and Reagents
4.2. Culturing of Human RASFs and THP1
4.3. Treatment of RASFs and THP-1
4.4. Assay for Cytokine Production
4.5. Western Immunoblotting
4.6. Cell Fractionation
4.7. DNA Binding Assay for STAT3
4.8. Molecular Dynamics (MD) Simulation Studies
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Residue | Takinib Interaction | Function |
---|---|---|
E638 | H-Bond | Amide H-Bond works with Q644 to stabilize ligand [24] |
Q644 | H-Bond | Stabilizes side chain H-bonds [25] |
Y640 | Hydrophobic | Modulates Tyr705 phosphorylation, pocket hydrophobicity [26] |
Y657 | H-bond, Hydrophobic | Works with E638 & Y640 to modulate pocket hydrophobicity [22,24,26] |
I659 | Hydrophobic | Ligand stabilization, pocket hydrophobicity [24,26] |
W623 | Hydrophobic, π-π | Induces ligand conformational fit, stabilizes ligand in pocket [24,27] |
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Panipinto, P.M.; Singh, A.K.; Shaikh, F.S.; Siegel, R.J.; Chourasia, M.; Ahmed, S. Takinib Inhibits Inflammation in Human Rheumatoid Arthritis Synovial Fibroblasts by Targeting the Janus Kinase-Signal Transducer and Activator of Transcription 3 (JAK/STAT3) Pathway. Int. J. Mol. Sci. 2021, 22, 12580. https://doi.org/10.3390/ijms222212580
Panipinto PM, Singh AK, Shaikh FS, Siegel RJ, Chourasia M, Ahmed S. Takinib Inhibits Inflammation in Human Rheumatoid Arthritis Synovial Fibroblasts by Targeting the Janus Kinase-Signal Transducer and Activator of Transcription 3 (JAK/STAT3) Pathway. International Journal of Molecular Sciences. 2021; 22(22):12580. https://doi.org/10.3390/ijms222212580
Chicago/Turabian StylePanipinto, Paul M., Anil K. Singh, Farheen S. Shaikh, Ruby J. Siegel, Mukesh Chourasia, and Salahuddin Ahmed. 2021. "Takinib Inhibits Inflammation in Human Rheumatoid Arthritis Synovial Fibroblasts by Targeting the Janus Kinase-Signal Transducer and Activator of Transcription 3 (JAK/STAT3) Pathway" International Journal of Molecular Sciences 22, no. 22: 12580. https://doi.org/10.3390/ijms222212580