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Title: Discovery of Peptidomimetic Ligands of EED as Allosteric Inhibitors of PRC2

Abstract

The function of EED within polycomb repressive complex 2 (PRC2) is mediated by a complex network of protein–protein interactions. Allosteric activation of PRC2 by binding of methylated proteins to the embryonic ectoderm development (EED) aromatic cage is essential for full catalytic activity, but details of this regulation are not fully understood. EED’s recognition of the product of PRC2 activity, histone H3 lysine 27 trimethylation (H3K27me3), stimulates PRC2 methyltransferase activity at adjacent nucleosomes leading to H3K27me3 propagation and, ultimately, gene repression. By coupling combinatorial chemistry and structure-based design, we optimized a low-affinity methylated jumonji, AT-rich interactive domain 2 (Jarid2) peptide to a smaller, more potent peptidomimetic ligand (Kd = 1.14 ± 0.14 μM) of the aromatic cage of EED. Our strategy illustrates the effectiveness of applying combinatorial chemistry to achieve both ligand potency and property optimization. Furthermore, the resulting ligands, UNC5114 and UNC5115, demonstrate that targeted disruption of EED’s reader function can lead to allosteric inhibition of PRC2 catalytic activity.

Authors:
 [1];  [2];  [1];  [1];  [1];  [2];  [1];  [2];  [2];  [2];  [1]; ORCiD logo [1]
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  1. Univ. of North Carolina, Chapel Hill, NC (United States)
  2. Univ. of Toronto, ON (Canada)
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1351384
Grant/Contract Number:  
R01GM100919
Resource Type:
Accepted Manuscript
Journal Name:
ACS Combinatorial Science
Additional Journal Information:
Journal Volume: 19; Journal Issue: 3; Journal ID: ISSN 2156-8952
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
ENGLISH
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 60 APPLIED LIFE SCIENCES; Peptides and proteins; Ligands; Monomers; Catalytic activity; Aromatic compounds

Citation Formats

Barnash, Kimberly D., The, Juliana, Norris-Drouin, Jacqueline L., Cholensky, Stephanie H., Worley, Beau M., Li, Fengling, Stuckey, Jacob I., Brown, Peter J., Vedadi, Masoud, Arrowsmith, Cheryl H., Frye, Stephen V., and James, Lindsey I. Discovery of Peptidomimetic Ligands of EED as Allosteric Inhibitors of PRC2. United States: N. p., 2017. Web. doi:10.1021/acscombsci.6b00174.
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Barnash, Kimberly D., The, Juliana, Norris-Drouin, Jacqueline L., Cholensky, Stephanie H., Worley, Beau M., Li, Fengling, Stuckey, Jacob I., Brown, Peter J., Vedadi, Masoud, Arrowsmith, Cheryl H., Frye, Stephen V., & James, Lindsey I. Discovery of Peptidomimetic Ligands of EED as Allosteric Inhibitors of PRC2. United States. https://doi.org/10.1021/acscombsci.6b00174
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Barnash, Kimberly D., The, Juliana, Norris-Drouin, Jacqueline L., Cholensky, Stephanie H., Worley, Beau M., Li, Fengling, Stuckey, Jacob I., Brown, Peter J., Vedadi, Masoud, Arrowsmith, Cheryl H., Frye, Stephen V., and James, Lindsey I. Wed . "Discovery of Peptidomimetic Ligands of EED as Allosteric Inhibitors of PRC2". United States. https://doi.org/10.1021/acscombsci.6b00174. https://www.osti.gov/servlets/purl/1351384.
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@article{osti_1351384,
title = {Discovery of Peptidomimetic Ligands of EED as Allosteric Inhibitors of PRC2},
author = {Barnash, Kimberly D. and The, Juliana and Norris-Drouin, Jacqueline L. and Cholensky, Stephanie H. and Worley, Beau M. and Li, Fengling and Stuckey, Jacob I. and Brown, Peter J. and Vedadi, Masoud and Arrowsmith, Cheryl H. and Frye, Stephen V. and James, Lindsey I.},
abstractNote = {The function of EED within polycomb repressive complex 2 (PRC2) is mediated by a complex network of protein–protein interactions. Allosteric activation of PRC2 by binding of methylated proteins to the embryonic ectoderm development (EED) aromatic cage is essential for full catalytic activity, but details of this regulation are not fully understood. EED’s recognition of the product of PRC2 activity, histone H3 lysine 27 trimethylation (H3K27me3), stimulates PRC2 methyltransferase activity at adjacent nucleosomes leading to H3K27me3 propagation and, ultimately, gene repression. By coupling combinatorial chemistry and structure-based design, we optimized a low-affinity methylated jumonji, AT-rich interactive domain 2 (Jarid2) peptide to a smaller, more potent peptidomimetic ligand (Kd = 1.14 ± 0.14 μM) of the aromatic cage of EED. Our strategy illustrates the effectiveness of applying combinatorial chemistry to achieve both ligand potency and property optimization. Furthermore, the resulting ligands, UNC5114 and UNC5115, demonstrate that targeted disruption of EED’s reader function can lead to allosteric inhibition of PRC2 catalytic activity.},
doi = {10.1021/acscombsci.6b00174},
journal = {ACS Combinatorial Science},
number = 3,
volume = 19,
place = {United States},
year = {Wed Feb 22 00:00:00 EST 2017},
month = {Wed Feb 22 00:00:00 EST 2017}
}
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Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1021/acscombsci.6b00174
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Cited by: 35 works
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Figures / Tables:

Figure 1 Figure 1: (a) Structure of truncated Jarid2 peptide, Jarid2114-118K116me3. This native ligand served as the scaffold for subsequent ligand optimization studies and soluble competitor for the first generation combinatorial library. (b) The library screening scheme is presented. The OBOC library is incubated with His-tagged Kme reader proteins, excess protein ismore » removed by rinsing. Protein G Dynabeads coated in anti-His antibody then enable magnetic enrichment of hit compounds while subsequent treatment with a soluble competitor elutes off lower affinity hits. (c) Jarid2114-118K116me3-derivative library was designed to probe ligand requirements at positions R1-R4. The native Jarid2 sequence was retained as the linker while also including methionine to enable hit cleavage via CNBr treatment. (d) High redundancy EED ligands. Both analogs were enriched via the magnetic enrichment assay with greater than two-fold redundancy« less
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    Getting a handle on chemical probes of chomatin readers
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    • Simhadri, Chakravarthi; Daze, Kevin D.; Douglas, Sarah F.
    • Universität des Saarlandes
    • DOI: 10.22028/d291-30928

    Discovery of selective activators of PRC2 mutant EED-I363M
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      Figure 3 (p. 14)figure
      Table 1 ; Part 1 (p. 16)table
      Table 1: Part 2 (p. 17)table
      Figure 4 (p. 19)figure
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