Genes to Cognition Programme, The Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, UK; KU Leuven, Center for Human Genetics and Leuven Institute for Neurodegenerative Diseases (LIND), and VIB Center for the Biology of Disease, Leuven, Belgium.
Proteomic Mass Spectrometry, The Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, UK.
Cell Rep. 2017 Oct 17;21(3):679-691. doi: 10.1016/j.celrep.2017.09.045.
Arc is an activity-regulated neuronal protein, but little is known about its interactions, assembly into multiprotein complexes, and role in human disease and cognition. We applied an integrated proteomic and genetic strategy by targeting a tandem affinity purification (TAP) tag and Venus fluorescent protein into the endogenous Arc gene in mice. This allowed biochemical and proteomic characterization of native complexes in wild-type and knockout mice. We identified many Arc-interacting proteins, of which PSD95 was the most abundant. PSD95 was essential for Arc assembly into 1.5-MDa complexes and activity-dependent recruitment to excitatory synapses. Integrating human genetic data with proteomic data showed that Arc-PSD95 complexes are enriched in schizophrenia, intellectual disability, autism, and epilepsy mutations and normal variants in intelligence. We propose that Arc-PSD95 postsynaptic complexes potentially affect human cognitive function.
Arc 是一种活性调节神经元蛋白，但人们对其相互作用、组装成多蛋白复合物以及在人类疾病和认知中的作用知之甚少。我们通过将串联亲和纯化 (TAP) 标签和 Venus 荧光蛋白靶向内源性 Arc 基因，应用了一种整合的蛋白质组学和遗传策略在小鼠中。这使得我们能够对野生型和敲除小鼠中的天然复合物进行生化和蛋白质组学分析。我们鉴定了许多与 Arc 相互作用的蛋白质，其中 PSD95 是最丰富的。PSD95 对于 Arc 组装成 1.5MDa 复合物以及活性依赖性募集到兴奋性突触是必不可少的。将人类遗传数据与蛋白质组学数据整合表明，Arc-PSD95 复合物在精神分裂症、智力障碍、自闭症和癫痫的突变以及智力正常变异中富集。我们提出，Arc-PSD95 突触后复合物可能会影响人类的认知功能。
