The oncoprotein YAP (Yes-associated protein) requires the TEAD family of transcription factors for the up-regulation of genes important for cell proliferation. Disrupting YAP–TEAD interaction is an attractive strategy for cancer therapy. Targeting TEADs using small molecules that either bind to the YAP-binding pocket or the palmitate-binding pocket is proposed to disrupt the YAP–TEAD interaction. There is a need for methodologies to facilitate robust and reliable identification of compounds that occupy either YAP-binding pocket or palmitate-binding pocket. Here, using NMR spectroscopy, we validated compounds that bind to these pockets and also identify the residues in mouse TEAD4 (mTEAD4) that interact with these compounds. Flufenamic acid (FA) was used as a positive control for validation of palmitate-binding pocket-occupying compounds by NMR. Furthermore, we identify a hit from a fragment screen and show that it occupies a site close to YAP-binding pocket on the TEAD surface. Our results also indicate that purified mTEAD4 can catalyze autopalmitoylation. NMR studies on mTEAD4 revealed that exchanges exist in TEAD as NMR signal broadening was observed for residues close to the palmitoylation site. Mutating the palmitoylated cysteine (C360S mutant) abolished palmitoylation, while no significant changes in the NMR spectrum were observed for the mutant which still binds to YAP. We also show that FA inhibits TEAD autopalmitoylation. Our studies highlight the utility of NMR spectroscopy in identifying small molecules that bind to TEAD pockets and reinforce the notion that both palmitate-binding pocket and YAP-binding pocket are targetable.
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June 2018
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A digital illustration of lung cancer cells. In this issue of the Biochemical Journal, Windhorst et al. investigate the effect of the actin- and calcium-regulating activities of ITPKB on the metastatic potential of lung cancer cells; for details, see pages 2057–2071.
Research Article|
June 26 2018
Structural and ligand-binding analysis of the YAP-binding domain of transcription factor TEAD4
Yan Li;
Yan Li
*
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Shuang Liu;
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Elizabeth Yihui Ng;
Elizabeth Yihui Ng
*
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Rong Li;
Rong Li
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Anders Poulsen;
Anders Poulsen
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Jeffrey Hill;
Jeffrey Hill
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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Ajaybabu V. Pobbati;
2The Cancer and Developmental Cell Biology Division, Institute of Molecular and Cell Biology, A*STAR, 61 Biopolis Drive, Singapore 138673
Correspondence: Ajaybabu V. Pobbati (ajaybabuvp@imcb.a-star.edu.sg), Alvin W. Hung (whung@etc.a-star.edu.sg) or CongBao Kang (cbkang@etc.a-star.edu.sg)
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Alvin W. Hung;
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
Correspondence: Ajaybabu V. Pobbati (ajaybabuvp@imcb.a-star.edu.sg), Alvin W. Hung (whung@etc.a-star.edu.sg) or CongBao Kang (cbkang@etc.a-star.edu.sg)
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Wanjin Hong
;
Wanjin Hong
2The Cancer and Developmental Cell Biology Division, Institute of Molecular and Cell Biology, A*STAR, 61 Biopolis Drive, Singapore 138673
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Thomas H. Keller;
Thomas H. Keller
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
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CongBao Kang
1Experimental Therapeutics Centre, A*STAR, 31 Biopolis Way, #3-01, Singapore 138669
Correspondence: Ajaybabu V. Pobbati (ajaybabuvp@imcb.a-star.edu.sg), Alvin W. Hung (whung@etc.a-star.edu.sg) or CongBao Kang (cbkang@etc.a-star.edu.sg)
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Publisher: Portland Press Ltd
Received:
March 17 2018
Revision Received:
May 07 2018
Accepted:
May 10 2018
Accepted Manuscript online:
May 14 2018
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society
2018
Biochem J (2018) 475 (12): 2043–2055.
Article history
Received:
March 17 2018
Revision Received:
May 07 2018
Accepted:
May 10 2018
Accepted Manuscript online:
May 14 2018
Citation
Yan Li, Shuang Liu, Elizabeth Yihui Ng, Rong Li, Anders Poulsen, Jeffrey Hill, Ajaybabu V. Pobbati, Alvin W. Hung, Wanjin Hong, Thomas H. Keller, CongBao Kang; Structural and ligand-binding analysis of the YAP-binding domain of transcription factor TEAD4. Biochem J 29 June 2018; 475 (12): 2043–2055. doi: https://doi.org/10.1042/BCJ20180225
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