Division of Biology, California Institute of Technology, Pasadena, CA, USA.
Proteomics. 2012 Aug;12(15-16):2464-76. doi: 10.1002/pmic.201200112.
Local protein synthesis and its activity-dependent modulation via dopamine receptor stimulation play an important role in synaptic plasticity - allowing synapses to respond dynamically to changes in their activity patterns. We describe here the metabolic labeling, enrichment, and MS-based identification of candidate proteins specifically translated in intact hippocampal neuropil sections upon treatment with the selective D1/D5 receptor agonist SKF81297. Using the noncanonical amino acid azidohomoalanine and click chemistry, we identified over 300 newly synthesized proteins specific to dendrites and axons. Candidates specific for the SKF81297-treated samples were predominantly involved in protein synthesis and synapse-specific functions. Furthermore, we demonstrate a dendrite-specific increase in proteins synthesis upon application of SKF81297. This study provides the first snapshot in the dynamics of the dopaminergic hippocampal neuropil proteome.
局部蛋白质合成及其活性依赖性调制通过多巴胺受体刺激在突触可塑性中起着重要作用——使突触能够对其活动模式的变化做出动态响应。我们在这里描述了代谢标记、富集和基于 MS 的鉴定方法，用于鉴定在使用选择性 D1/D5 受体激动剂 SKF81297 处理完整海马神经突切片时特异性翻译的候选蛋白。我们使用非典型氨基酸叠氮基高丙氨酸和点击化学，鉴定了超过 300 种新合成的蛋白质，这些蛋白质特异性存在于树突和轴突中。候选物特异性针对 SKF81297 处理的样本主要涉及蛋白质合成和突触特异性功能。此外，我们证明了 SKF81297 应用后蛋白质合成的树突特异性增加。这项研究提供了多巴胺能海马神经突蛋白质组动态的第一个快照。
