Selectivity via Cooperativity: Preferential Stabilization of the p65/14-3-3 Interaction with Semisynthetic Natural Products

Madita Wolter; Pim De Vink; João Filipe Neves; Sonja Srdanović; Yusuke Higuchi; Nobuo Kato; Andrew Wilson; Isabelle Landrieu; Luc Brunsveld; Christian Ottmann

doi:10.1021/jacs.0c02151

Selectivity via Cooperativity: Preferential Stabilization of the p65/14-3-3 Interaction with Semisynthetic Natural Products

Madita Wolter, Pim De Vink, João Filipe Neves, Sonja Srdanović, Yusuke Higuchi, Nobuo Kato, Andrew Wilson, Isabelle Landrieu, Luc Brunsveld, Christian Ottmann^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

31 Citations (Scopus)

Abstract

Natural compounds are an important class of potent drug molecules including some retrospectively found to act as stabilizers of protein-protein interactions (PPIs). However, the design of synthetic PPI stabilizers remains an understudied approach. To date, there are limited examples where cooperativity has been utilized to guide the optimization of a PPI stabilizer. The 14-3-3 scaffold proteins provide an excellent platform to explore PPI stabilization because these proteins mediate several hundred PPIs, and a class of natural compounds, the fusicoccanes, are known to stabilize a subset of 14-3-3 protein interactions. 14-3-3 has been reported to negatively regulate the p65 subunit of the NF-κB transcription factor, which qualifies this protein complex as a potential target for drug discovery to control cell proliferation. Here, we report the high-resolution crystal structures of two 14-3-3 binding motifs of p65 in complex with 14-3-3. A semisynthetic natural product derivative, DP-005, binds to an interface pocket of the p65/14-3-3 complex and concomitantly stabilizes it. Cooperativity analyses of this interaction, and other disease relevant 14-3-3-PPIs, demonstrated selectivity of DP-005 for the p65/14-3-3 complex. The adaptation of a cooperative binding model provided a general approach to characterize stabilization and to assay for selectivity of PPI stabilizers.

Original language	English
Pages (from-to)	11772-11783
Number of pages	12
Journal	Journal of the American Chemical Society
Volume	142
Issue number	27
DOIs	https://doi.org/10.1021/jacs.0c02151
Publication status	Published - 8 Jul 2020

Bibliographical note

Funding Information:
We thank François-Xavier Cantrelle from the University of Lille for NMR data acquisition. This research is supported by funding from the European Union through the TASPPI project (H2020-MSCA-ITN-2015, grant number 675179) and by the LabEx (Laboratory of Excellence) DISTALZ (ANR, ANR-11-LABX-009) and through The Netherlands Organization for Scientific Research (NWO) via VICI grant 016.150.366 and via Gravity Program 024.001.035. The NMR facilities were funded by the Nord Region Council, CNRS, Institut Pasteur de Lille, the European Community (ERDF), the French Ministry of Research, and the University of Lille. We acknowledge support for the NMR facilities from TGE RMN THC (CNRS, FR-3050) and FRABio (Univ. Lille, CNRS, FR-3688). This work was supported by the EPSRC (EP/N013573/1) and The Wellcome Trust (097827/Z/11/A).

Publisher Copyright:
© 2020 American Chemical Society.

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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@article{2681138b5d3746c7a07875ee4b40d52f,

title = "Selectivity via Cooperativity: Preferential Stabilization of the p65/14-3-3 Interaction with Semisynthetic Natural Products",

abstract = "Natural compounds are an important class of potent drug molecules including some retrospectively found to act as stabilizers of protein-protein interactions (PPIs). However, the design of synthetic PPI stabilizers remains an understudied approach. To date, there are limited examples where cooperativity has been utilized to guide the optimization of a PPI stabilizer. The 14-3-3 scaffold proteins provide an excellent platform to explore PPI stabilization because these proteins mediate several hundred PPIs, and a class of natural compounds, the fusicoccanes, are known to stabilize a subset of 14-3-3 protein interactions. 14-3-3 has been reported to negatively regulate the p65 subunit of the NF-κB transcription factor, which qualifies this protein complex as a potential target for drug discovery to control cell proliferation. Here, we report the high-resolution crystal structures of two 14-3-3 binding motifs of p65 in complex with 14-3-3. A semisynthetic natural product derivative, DP-005, binds to an interface pocket of the p65/14-3-3 complex and concomitantly stabilizes it. Cooperativity analyses of this interaction, and other disease relevant 14-3-3-PPIs, demonstrated selectivity of DP-005 for the p65/14-3-3 complex. The adaptation of a cooperative binding model provided a general approach to characterize stabilization and to assay for selectivity of PPI stabilizers.",

author = "Madita Wolter and {De Vink}, Pim and Neves, {Jo{\~a}o Filipe} and Sonja Srdanovi{\'c} and Yusuke Higuchi and Nobuo Kato and Andrew Wilson and Isabelle Landrieu and Luc Brunsveld and Christian Ottmann",

note = "Funding Information: We thank Fran{\c c}ois-Xavier Cantrelle from the University of Lille for NMR data acquisition. This research is supported by funding from the European Union through the TASPPI project (H2020-MSCA-ITN-2015, grant number 675179) and by the LabEx (Laboratory of Excellence) DISTALZ (ANR, ANR-11-LABX-009) and through The Netherlands Organization for Scientific Research (NWO) via VICI grant 016.150.366 and via Gravity Program 024.001.035. The NMR facilities were funded by the Nord Region Council, CNRS, Institut Pasteur de Lille, the European Community (ERDF), the French Ministry of Research, and the University of Lille. We acknowledge support for the NMR facilities from TGE RMN THC (CNRS, FR-3050) and FRABio (Univ. Lille, CNRS, FR-3688). This work was supported by the EPSRC (EP/N013573/1) and The Wellcome Trust (097827/Z/11/A). Publisher Copyright: {\textcopyright} 2020 American Chemical Society.",

year = "2020",

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doi = "10.1021/jacs.0c02151",

language = "English",

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AU - Wolter, Madita

AU - De Vink, Pim

AU - Neves, João Filipe

AU - Srdanović, Sonja

AU - Higuchi, Yusuke

AU - Kato, Nobuo

AU - Wilson, Andrew

AU - Landrieu, Isabelle

AU - Brunsveld, Luc

AU - Ottmann, Christian

N1 - Funding Information: We thank François-Xavier Cantrelle from the University of Lille for NMR data acquisition. This research is supported by funding from the European Union through the TASPPI project (H2020-MSCA-ITN-2015, grant number 675179) and by the LabEx (Laboratory of Excellence) DISTALZ (ANR, ANR-11-LABX-009) and through The Netherlands Organization for Scientific Research (NWO) via VICI grant 016.150.366 and via Gravity Program 024.001.035. The NMR facilities were funded by the Nord Region Council, CNRS, Institut Pasteur de Lille, the European Community (ERDF), the French Ministry of Research, and the University of Lille. We acknowledge support for the NMR facilities from TGE RMN THC (CNRS, FR-3050) and FRABio (Univ. Lille, CNRS, FR-3688). This work was supported by the EPSRC (EP/N013573/1) and The Wellcome Trust (097827/Z/11/A). Publisher Copyright: © 2020 American Chemical Society.

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N2 - Natural compounds are an important class of potent drug molecules including some retrospectively found to act as stabilizers of protein-protein interactions (PPIs). However, the design of synthetic PPI stabilizers remains an understudied approach. To date, there are limited examples where cooperativity has been utilized to guide the optimization of a PPI stabilizer. The 14-3-3 scaffold proteins provide an excellent platform to explore PPI stabilization because these proteins mediate several hundred PPIs, and a class of natural compounds, the fusicoccanes, are known to stabilize a subset of 14-3-3 protein interactions. 14-3-3 has been reported to negatively regulate the p65 subunit of the NF-κB transcription factor, which qualifies this protein complex as a potential target for drug discovery to control cell proliferation. Here, we report the high-resolution crystal structures of two 14-3-3 binding motifs of p65 in complex with 14-3-3. A semisynthetic natural product derivative, DP-005, binds to an interface pocket of the p65/14-3-3 complex and concomitantly stabilizes it. Cooperativity analyses of this interaction, and other disease relevant 14-3-3-PPIs, demonstrated selectivity of DP-005 for the p65/14-3-3 complex. The adaptation of a cooperative binding model provided a general approach to characterize stabilization and to assay for selectivity of PPI stabilizers.

AB - Natural compounds are an important class of potent drug molecules including some retrospectively found to act as stabilizers of protein-protein interactions (PPIs). However, the design of synthetic PPI stabilizers remains an understudied approach. To date, there are limited examples where cooperativity has been utilized to guide the optimization of a PPI stabilizer. The 14-3-3 scaffold proteins provide an excellent platform to explore PPI stabilization because these proteins mediate several hundred PPIs, and a class of natural compounds, the fusicoccanes, are known to stabilize a subset of 14-3-3 protein interactions. 14-3-3 has been reported to negatively regulate the p65 subunit of the NF-κB transcription factor, which qualifies this protein complex as a potential target for drug discovery to control cell proliferation. Here, we report the high-resolution crystal structures of two 14-3-3 binding motifs of p65 in complex with 14-3-3. A semisynthetic natural product derivative, DP-005, binds to an interface pocket of the p65/14-3-3 complex and concomitantly stabilizes it. Cooperativity analyses of this interaction, and other disease relevant 14-3-3-PPIs, demonstrated selectivity of DP-005 for the p65/14-3-3 complex. The adaptation of a cooperative binding model provided a general approach to characterize stabilization and to assay for selectivity of PPI stabilizers.

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Selectivity via Cooperativity: Preferential Stabilization of the p65/14-3-3 Interaction with Semisynthetic Natural Products

Abstract

Bibliographical note

ASJC Scopus subject areas

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