SNAP-25 is a target of protein kinase C phosphorylation critical to NMDA receptor trafficking.
SNAP-25 is a target of protein kinase C phosphorylation critical to NMDA receptor trafficking.
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DOI：
10.1523/jneurosci.4933-08.2010
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发表时间：
2010-01-06
期刊：
影响因子：
--
通讯作者：
Zukin RS
中科院分区：
文献类型：
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作者：
Lau CG;Takayasu Y;Rodenas-Ruano A;Paternain AV;Lerma J;Bennett MV;Zukin RS
蛋白激酶C（PKC）增强NMDA受体（NMDAR）通道开放速率，并通过SNARE依赖性胞吐作用促进NMDAR递送至细胞表面。虽然PKC增强的机制已经建立，但PKC的分子靶点尚不清楚。在这里，我们证明了25 kDa突触体相关蛋白（SNAP-25）（一种SNARE蛋白）在功能上与PKC依赖性NMDAR插入相关，并确定丝氨酸残基-187为PKC磷酸化的分子靶点。组成性活性PKC通过贴片移液管传递增强海马神经元的NMDA（但不是AMPA）全细胞电流。靶向SNAP-25或突变体SNAP-25（S187 A）的RNAi的表达和/或通过用BoNT A、BoNT B或SNAP-25 C-末端封闭肽处理的SNARE复合物的急性破坏消除了NMDAR增强。SNAP-25肽和功能阻断抗体抑制苔藓纤维-CA 3突触处NMDA EPSC的PKC增强。这些研究结果确定SNAP-25作为PKC磷酸化的目标，PKC依赖性纳入突触NMDAR的关键，并记录了突触可塑性相关的主要SNARE蛋白的突触后作用。
Protein kinase C (PKC) enhances NMDA receptor (NMDAR) channel opening rate and promotes NMDAR delivery to the cell surface via SNARE-dependent exocytosis. Although the mechanisms of PKC potentiation are established, the molecular target of PKC is unclear. Here we show that synaptosomal-associated protein of 25 kDa (SNAP-25), a SNARE protein, is functionally relevant to PKC-dependent NMDAR insertion and identify serine residue-187 as the molecular target of PKC phosphorylation. Constitutively active PKC delivered via the patch pipette potentiated NMDA (but not AMPA) whole-cell currents in hippocampal neurons. Expression of RNAi targeting SNAP-25 or mutant SNAP-25(S187A) and/or acute disruption of the SNARE complex by treatment with BoNT A, BoNT B or SNAP-25 C-terminal blocking peptide abolished NMDAR potentiation. A SNAP-25 peptide and function-blocking antibody suppressed PKC potentiation of NMDA EPSCs at mossy fiber-CA3 synapses. These findings identify SNAP-25 as the target of PKC phosphorylation critical to PKC-dependent incorporation of synaptic NMDARs and document a postsynaptic action of this major SNARE protein relevant to synaptic plasticity.