TY - JOUR
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 -