Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD

Thor S. Thorsen, Kenneth L. Madsen, Nelson Rebola, Mette Rathje, Victor Anggono, Anders Bach, Irina S. Moreira, Nicolai Stuhr-Hansen, Tino Dyhring, Dan Peters, Thijs Beuming, Richard L Huganir, Harel Weinstein, Christophe Mulle, Kristian Strømgaard, Lars Christian B Rønn, Ulrik Gether

Research output: Contribution to journalArticle

Abstract

Proteins containing PSD-95/Discs-large/ZO-1 homology (PDZ) domains play key roles in the assembly and regulation of cellular signaling pathways and represent putative targets for new pharmacotherapeutics. Here we describe the first small-molecule inhibitor (FSC231) of the PDZdomain inprotein interacting with C kinase 1 (PICK1) identified by a screening of ∼44,000 compounds in a fluorescent polarization assay. The inhibitor bound the PICK1 PDZ domain with an affinity similar to that observed for endogenous peptide ligands (K i∼10.1 μM). Mutational analysis, together with computational docking of the compound in simulations starting from the PDZ domain structure, identified the binding mode of FSC231. The specificityof FSC231 for the PICK1 PDZ domain was supported by the lack of binding to PDZ domains of postsynaptic density protein 95 (PSD-95) and glutamate receptor interacting protein 1 (GRIP1). Pretreatment of cultured hippocampal neurons with FSC231 inhibited coimmunopreciptation of the AMPA receptor GluR2 subunit with PICK1. In agreement with inhibiting the role of PICK1 in GluR2 trafficking, FSC231 accelerated recycling of pHluorin-tagged GluR2 in hippocampal neurons after internalization in response to NMDA receptor activation. FSC231 blocked the expression of both long-term depression and long-term potentiation in hippocampal CA1 neurons from acute slices, consistent with inhibition of the bidirectional function of PICK1 in synaptic plasticity. Given the proposedrole of the PICK1/AMPA receptor interaction in neuropathic pain, excitotoxicity, and cocaine addiction, FSC231 might serve as a lead in the future development of new therapeutics against these conditions.

Original languageEnglish (US)
Pages (from-to)413-418
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number1
DOIs
StatePublished - 2010

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PDZ Domains
Phosphotransferases
Neurons
Receptor-Interacting Protein Serine-Threonine Kinases
Cocaine-Related Disorders
Neuronal Plasticity
AMPA Receptors
Long-Term Potentiation
Glutamate Receptors
Recycling
Neuralgia
FSC 231
N-Methyl-D-Aspartate Receptors
Depression
Ligands
Peptides

Keywords

  • AMPA receptors
  • Drug discovery
  • Fluorescence polarization
  • Protein-protein interactions
  • Synaptic plasticity

ASJC Scopus subject areas

  • General

Cite this

Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD. / Thorsen, Thor S.; Madsen, Kenneth L.; Rebola, Nelson; Rathje, Mette; Anggono, Victor; Bach, Anders; Moreira, Irina S.; Stuhr-Hansen, Nicolai; Dyhring, Tino; Peters, Dan; Beuming, Thijs; Huganir, Richard L; Weinstein, Harel; Mulle, Christophe; Strømgaard, Kristian; Rønn, Lars Christian B; Gether, Ulrik.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 1, 2010, p. 413-418.

Research output: Contribution to journalArticle

Thorsen, TS, Madsen, KL, Rebola, N, Rathje, M, Anggono, V, Bach, A, Moreira, IS, Stuhr-Hansen, N, Dyhring, T, Peters, D, Beuming, T, Huganir, RL, Weinstein, H, Mulle, C, Strømgaard, K, Rønn, LCB & Gether, U 2010, 'Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD', Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 1, pp. 413-418. https://doi.org/10.1073/pnas.0902225107
Thorsen, Thor S. ; Madsen, Kenneth L. ; Rebola, Nelson ; Rathje, Mette ; Anggono, Victor ; Bach, Anders ; Moreira, Irina S. ; Stuhr-Hansen, Nicolai ; Dyhring, Tino ; Peters, Dan ; Beuming, Thijs ; Huganir, Richard L ; Weinstein, Harel ; Mulle, Christophe ; Strømgaard, Kristian ; Rønn, Lars Christian B ; Gether, Ulrik. / Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 1. pp. 413-418.
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T1 - Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD

AU - Thorsen, Thor S.

AU - Madsen, Kenneth L.

AU - Rebola, Nelson

AU - Rathje, Mette

AU - Anggono, Victor

AU - Bach, Anders

AU - Moreira, Irina S.

AU - Stuhr-Hansen, Nicolai

AU - Dyhring, Tino

AU - Peters, Dan

AU - Beuming, Thijs

AU - Huganir, Richard L

AU - Weinstein, Harel

AU - Mulle, Christophe

AU - Strømgaard, Kristian

AU - Rønn, Lars Christian B

AU - Gether, Ulrik

PY - 2010

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N2 - Proteins containing PSD-95/Discs-large/ZO-1 homology (PDZ) domains play key roles in the assembly and regulation of cellular signaling pathways and represent putative targets for new pharmacotherapeutics. Here we describe the first small-molecule inhibitor (FSC231) of the PDZdomain inprotein interacting with C kinase 1 (PICK1) identified by a screening of ∼44,000 compounds in a fluorescent polarization assay. The inhibitor bound the PICK1 PDZ domain with an affinity similar to that observed for endogenous peptide ligands (K i∼10.1 μM). Mutational analysis, together with computational docking of the compound in simulations starting from the PDZ domain structure, identified the binding mode of FSC231. The specificityof FSC231 for the PICK1 PDZ domain was supported by the lack of binding to PDZ domains of postsynaptic density protein 95 (PSD-95) and glutamate receptor interacting protein 1 (GRIP1). Pretreatment of cultured hippocampal neurons with FSC231 inhibited coimmunopreciptation of the AMPA receptor GluR2 subunit with PICK1. In agreement with inhibiting the role of PICK1 in GluR2 trafficking, FSC231 accelerated recycling of pHluorin-tagged GluR2 in hippocampal neurons after internalization in response to NMDA receptor activation. FSC231 blocked the expression of both long-term depression and long-term potentiation in hippocampal CA1 neurons from acute slices, consistent with inhibition of the bidirectional function of PICK1 in synaptic plasticity. Given the proposedrole of the PICK1/AMPA receptor interaction in neuropathic pain, excitotoxicity, and cocaine addiction, FSC231 might serve as a lead in the future development of new therapeutics against these conditions.

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