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 Huganir; Harel Weinstein; Christophe Mulle; Kristian Strømgaard; Lars Christian B Rønn; Ulrik Gether

doi:10.1073/pnas.0902225107

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 Huganir, Harel Weinstein, Christophe Mulle, Kristian Strømgaard, Lars Christian B Rønn, Ulrik Gether

86 Citations (Scopus)

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 language	English
Journal	Proceedings of the National Academy of Science of the United States of America
Volume	107
Issue number	1
Pages (from-to)	413-8
Number of pages	5
ISSN	0027-8424
DOIs	https://doi.org/10.1073/pnas.0902225107
Publication status	Published - 2010

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10.1073/pnas.0902225107

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Thorsen, T. S., Madsen, K. L., Rebola, N., Rathje, M., Anggono, V., Bach, A., Moreira, I. S., Stuhr-Hansen, N., Dyhring, T., Peters, D., Beuming, T., Huganir, R., Weinstein, H., Mulle, C., Strømgaard, K., Rønn, L. C. B., & 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 Science of the United States of America, 107(1), 413-8. https://doi.org/10.1073/pnas.0902225107

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 et al.
In: Proceedings of the National Academy of Science of the United States of America, Vol. 107, No. 1, 2010, p. 413-8.

Research output: Contribution to journal › Journal article › Research › peer-review

Thorsen, TS , Madsen, KL, Rebola, N, Rathje, M, Anggono, V, Bach, A, Moreira, IS, Stuhr-Hansen, N, Dyhring, T, Peters, D, Beuming, T, Huganir, R, 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 Science of the United States of America, vol. 107, no. 1, pp. 413-8. https://doi.org/10.1073/pnas.0902225107

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

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.",

author = "Thorsen, {Thor S} and Madsen, {Kenneth L} and Nelson Rebola and Mette Rathje and Victor Anggono and Anders Bach and Moreira, {Irina S} and Nicolai Stuhr-Hansen and Tino Dyhring and Dan Peters and Thijs Beuming and Richard Huganir and Harel Weinstein and Christophe Mulle and Kristian Str{\o}mgaard and R{\o}nn, {Lars Christian B} and Ulrik Gether",

note = "Keywords: Animals; Binding Sites; COS Cells; Carbamates; Carrier Proteins; Cercopithecus aethiops; Cinnamates; Hippocampus; Humans; Long-Term Potentiation; Long-Term Synaptic Depression; Models, Molecular; Molecular Structure; Neuronal Plasticity; Neurons; Nuclear Proteins; PDZ Domains; Peptides; Protein Structure, Tertiary; Receptors, AMPA; Recombinant Fusion Proteins",

year = "2010",

doi = "10.1073/pnas.0902225107",

language = "English",

volume = "107",

pages = "413--8",

journal = "Proceedings of the National Academy of Science of the United States of America",

issn = "0027-8424",

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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

AU - Weinstein, Harel

AU - Mulle, Christophe

AU - Strømgaard, Kristian

AU - Rønn, Lars Christian B

AU - Gether, Ulrik

N1 - Keywords: Animals; Binding Sites; COS Cells; Carbamates; Carrier Proteins; Cercopithecus aethiops; Cinnamates; Hippocampus; Humans; Long-Term Potentiation; Long-Term Synaptic Depression; Models, Molecular; Molecular Structure; Neuronal Plasticity; Neurons; Nuclear Proteins; PDZ Domains; Peptides; Protein Structure, Tertiary; Receptors, AMPA; Recombinant Fusion Proteins

PY - 2010

Y1 - 2010

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.

AB - 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.

U2 - 10.1073/pnas.0902225107

DO - 10.1073/pnas.0902225107

M3 - Journal article

C2 - 20018661

SN - 0027-8424

VL - 107

SP - 413

EP - 418

JO - Proceedings of the National Academy of Science of the United States of America

JF - Proceedings of the National Academy of Science of the United States of America

IS - 1

ER -

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

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