Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD
Identification of a small-molecule inhibitor of the PICK1 PDZ domain that inhibits hippocampal LTP and LTD
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DOI：
10.1073/pnas.0902225107
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发表时间：
2010-01-05
影响因子：
11.1
通讯作者：
Gether, Ulrik
中科院分区：
文献类型：
--
作者：
Thorsen, Thor S.;Madsen, Kenneth L.;Gether, Ulrik
含有 PSD-95/Discs-large/ZO-1 同源 (PDZ) 结构域的蛋白质在细胞信号通路的组装和调节中发挥着关键作用，并代表了新药物治疗的假定靶点。在这里，我们描述了第一个与 C 激酶 1 (PICK1) 相互作用的蛋白质中 PDZ 结构域的小分子抑制剂 (FSC231)，该抑制剂是通过在荧光偏振测定中筛选类似的 44,000 种化合物而确定的。该抑制剂结合 PICK1 PDZ 结构域的亲和力与内源肽配体观察到的亲和力相似（K-i 类似于 10.1 μM）。突变分析以及从 PDZ 结构域结构开始的模拟中的化合物计算对接，确定了 FSC231 的结合模式。 FSC231 对 PICK1 PDZ 结构域的特异性得到了与突触后密度蛋白 95 (PSD-95) 和谷氨酸受体相互作用蛋白 1 (GRIP1) PDZ 结构域缺乏结合的支持。用 FSC231 预处理培养的海马神经元可抑制 AMPA 受体 GluR2 亚基与 PICK1 的免疫共沉淀。与抑制 PICK1 在 GluR2 运输中的作用相一致，FSC231 响应 NMDA 受体激活而内化后，加速了海马神经元中 pHluorin 标记的 GluR2 的回收。 FSC231 阻断急性切片海马 CA1 神经元中长时程抑制和长时程增强的表达，这与抑制 PICK1 在突触可塑性中的双向功能一致。鉴于 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作用，FSC231 可能成为未来开发针对这些疾病的新疗法的先导。
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 PDZ domain in protein interacting with C kinase 1(PICK1) identified by a screening of similar to 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 similar to 10.1 mu M). Mutational 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.