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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD
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Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD

机译:鉴定抑制海马LTP和LTD的PICK1 PDZ域小分子抑制剂

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摘要

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 pharmacotherapeu-tics. Here we describe the first small-molecule inhibitor (FSC231) of the PDZ domain in protein 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). Mutation-al analysis, together with computational docking of the compound in simulations starting from the PDZ domain structure, identified the binding mode of FSC231. The specificity of 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 proposed role 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.
机译:包含PSD-95 / Discs-large / ZO-1同源性(PDZ)域的蛋白质在细胞信号传导途径的组装和调节中起着关键作用,并代表了新药物治疗的假定靶标。在这里,我们描述了与C激酶1(PICK1)相互作用的蛋白质中PDZ域的第一个小分子抑制剂(FSC231),通过荧光偏振分析筛选了约44,000种化合物而鉴定出。该抑制剂以与内源肽配体所观察到的相似的亲和力(K_i〜10.1μM)结合PICK1 PDZ域。突变分析以及从PDZ域结构开始的模拟中化合物的计算对接,确定了FSC231的结合模式。缺乏与突触后密度蛋白95(PSD-95)和谷氨酸受体相互作用蛋白1(GRIP1)的PDZ结构域的结合,支持了FSC231对PICK1 PDZ结构域的特异性。用FSC231预处理培养的海马神经元可抑制AMICK受体GluR2亚基与PICK1的共免疫沉淀。与抑制PICK1在GluR2转运中的作用一致,FSC231在响应NMDA受体激活后,内化后在海马神经元中加速了pHluorin标记的GluR2的再循环。 FSC231阻断了急性切片海马CA1神经元中长期抑制和长期增强的表达,这与抑制PICK1在突触可塑性中的双向功能一致。考虑到PICK1 / AMPA受体相互作用在神经性疼痛,兴奋性毒性和可卡因成瘾中的拟议作用,FSC231可能会在未来开发针对这些疾病的新疗法中发挥主导作用。

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  • 作者单位

    Molecular Neuropharmacology Group and Center for Pharmacogenomics, Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark;

    Molecular Neuropharmacology Group and Center for Pharmacogenomics, Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark;

    Laboratoire Physiologie Cellulaire de la Synapse, CNRS UMR 5091, Bordeaux Neuroscience Institute, University of Bordeaux, 33077 Bordeaux, France;

    Molecular Neuropharmacology Group and Center for Pharmacogenomics, Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark;

    Department of Neuroscience, The Howard Hughes Medical Institute, The Johns Hopkins University School of Medicine, Baltimore, MD 21205;

    Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark;

    Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, NY 10021;

    Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark;

    Drug Discovery, Neurosearch A/S, DK-2750 Ballerup, Denmark;

    Drug Discovery, Neurosearch A/S, DK-2750 Ballerup, Denmark;

    Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, NY 10021;

    Department of Neuroscience, The Howard Hughes Medical Institute, The Johns Hopkins University School of Medicine, Baltimore, MD 21205;

    Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, NY 10021;

    Laboratoire Physiologie Cellulaire de la Synapse, CNRS UMR 5091, Bordeaux Neuroscience Institute, University of Bordeaux, 33077 Bordeaux, France;

    Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark;

    Drug Discovery, Neurosearch A/S, DK-2750 Ballerup, Denmark;

    Molecular Neuropharmacology Group and Center for Pharmacogenomics, Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    drug discovery; fluorescence polarization; protein-protein interactions; synaptic plasticity; AMPA receptors;

    机译:药物发现;荧光偏振蛋白质相互作用突触可塑性AMPA受体;

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