受体酪氨酸激酶MET相互作用组与发育中突触处的神经发育障碍相关蛋白
Receptor Tyrosine Kinase MET Interactome and Neurodevelopmental Disorder Partners at the Developing Synapse.
作者信息
Xie Zhihui, Li Jing, Baker Jonathan, Eagleson Kathie L, Coba Marcelo P, Levitt Pat
机构信息
Department of Pediatrics and The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California.
Zilkha Neurogenetic Institute, Keck School of Medicine of the University of Southern California, Los Angeles, California.
出版信息
Biol Psychiatry. 2016 Dec 15;80(12):933-942. doi: 10.1016/j.biopsych.2016.02.022. Epub 2016 Feb 26.
BACKGROUND
Atypical synapse development and plasticity are implicated in many neurodevelopmental disorders (NDDs). NDD-associated, high-confidence risk genes have been identified, yet little is known about functional relationships at the level of protein-protein interactions, which are the dominant molecular bases responsible for mediating circuit development.
METHODS
Proteomics in three independent developing neocortical synaptosomal preparations identified putative interacting proteins of the ligand-activated MET receptor tyrosine kinase, an autism risk gene that mediates synapse development. The candidates were translated into interactome networks and analyzed bioinformatically. Additionally, three independent quantitative proximity ligation assays in cultured neurons and four independent immunoprecipitation analyses of synaptosomes validated protein interactions.
RESULTS
Approximately 11% (8/72) of MET-interacting proteins, including SHANK3, SYNGAP1, and GRIN2B, are associated with NDDs. Proteins in the MET interactome were translated into a novel MET interactome network based on human protein-protein interaction databases. High-confidence genes from different NDD datasets that encode synaptosomal proteins were analyzed for being enriched in MET interactome proteins. This was found for autism but not schizophrenia, bipolar disorder, major depressive disorder, or attention-deficit/hyperactivity disorder. There is correlated gene expression between MET and its interactive partners in developing human temporal and visual neocortices but not with highly expressed genes that are not in the interactome. Proximity ligation assays and biochemical analyses demonstrate that MET-protein partner interactions are dynamically regulated by receptor activation.
CONCLUSIONS
The results provide a novel molecular framework for deciphering the functional relations of key regulators of synaptogenesis that contribute to both typical cortical development and to NDDs.
背景
非典型突触发育和可塑性与许多神经发育障碍（NDDs）有关。已鉴定出与NDD相关的高置信度风险基因，但对于蛋白质-蛋白质相互作用水平上的功能关系知之甚少，而蛋白质-蛋白质相互作用是介导神经回路发育的主要分子基础。
方法
对三种独立的发育中的新皮质突触体标本进行蛋白质组学分析，确定了配体激活的MET受体酪氨酸激酶的推定相互作用蛋白，MET是一种介导突触发育的自闭症风险基因。将候选蛋白转化为相互作用组网络并进行生物信息学分析。此外，在培养的神经元中进行了三项独立的定量邻近连接测定，以及对突触体进行了四项独立的免疫沉淀分析，以验证蛋白质相互作用。
结果
与MET相互作用的蛋白质中约11%（8/72），包括SHANK3、SYNGAP1和GRIN2B，与NDDs相关。基于人类蛋白质-蛋白质相互作用数据库，将MET相互作用组中的蛋白质转化为一个新的MET相互作用组网络。分析了来自不同NDD数据集的编码突触体蛋白的高置信度基因在MET相互作用组蛋白中的富集情况。在自闭症中发现了这种富集，但在精神分裂症、双相情感障碍、重度抑郁症或注意力缺陷多动障碍中未发现。在发育中的人类颞叶和视觉新皮质中，MET与其相互作用伙伴之间存在相关基因表达，但与相互作用组中未高度表达的基因不存在相关基因表达。邻近连接测定和生化分析表明，MET-蛋白质伙伴相互作用受受体激活动态调节。
结论
这些结果为解读突触发生关键调节因子的功能关系提供了一个新的分子框架，这些调节因子对典型皮质发育和NDDs都有贡献。
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