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Immunoprecipitation and mass spectrometry define TET1 interactome during oligodendrocyte differentiation
Cell and Bioscience ( IF 6.1 ) Pub Date : 2020-09-16 , DOI: 10.1186/s13578-020-00473-5
Ming Zhang , Kaixiang Zhang , Jian Wang , Yuming Liu , Guangxin Liu , Weilin Jin , Shengxi Wu , Xianghui Zhao

Ten-eleven translocation (TET) proteins, encoding dioxygenase for DNA hydroxymethylation, are important players in nervous system development and disease. In addition to their proverbial enzymatic role, TET proteins also possess non-enzymatic activity and function in multiple protein–protein interaction networks, which remains largely unknown during oligodendrocyte differentiation. To identify partners of TET1 in the myelinating cells, we performed proteome-wide analysis using co-immunoprecipitation coupled to mass spectrometry (IP-MS) in purified oligodendrocyte precursor cells (OPCs) and mature oligodendrocytes (mOLs), respectively. Following a stringent selection of MS data based on identification reliability and protein enrichment, we identified a core set of 1211 partners that specifically interact with TET1 within OPCs and OLs. Analysis of the biological process and pathways associated with TET1-interacting proteins indicates a significant enrichment of proteins involved in regulation of cellular protein localization, cofactor metabolic process and regulation of catabolic process, et al. We further validated TET1 interactions with selected partners. Overall, this comprehensive analysis of the endogenous TET1 interactome during oligodendrocyte differentiation suggest its novel mechanism in regulating oligodendrocyte homeostasis and provide comprehensive insight into the molecular pathways associated with TET1.

中文翻译:

免疫沉淀和质谱法定义少突胶质细胞分化过程中的TET1相互作用

编码DNA羟甲基化的双加氧酶的十一个11易位(TET)蛋白是神经系统发育和疾病的重要参与者。除其众所周知的酶作用外,TET蛋白还具有非酶活性,并在多种蛋白-蛋白相互作用网络中发挥作用,在少突胶质细胞分化过程中仍然未知。为了鉴定髓鞘细胞中TET1的伴侣,我们分别在纯化的少突胶质细胞前体细胞(OPC)和成熟少突胶质细胞(mOL)中使用共免疫沉淀与质谱(IP-MS)进行了蛋白质组分析。根据鉴定的可靠性和蛋白质富集对MS数据进行严格选择之后,我们确定了1211个核心伴侣的核心集合,这些伙伴与OPC和OL中的TET1特异性相互作用。与TET1相互作用蛋白相关的生物学过程和途径的分析表明,参与细胞蛋白定位,辅因子代谢过程和分解代谢过程调节等过程的蛋白质大量富集。我们进一步验证了与选定合作伙伴的TET1互动。总体而言,对少突胶质细胞分化过程中内源性TET1相互作用组的这一综合分析表明,其在调节少突胶质细胞稳态中的新机制,并为与TET1相关的分子途径提供了全面的见解。我们进一步验证了与选定合作伙伴的TET1互动。总体而言,对少突胶质细胞分化过程中内源性TET1相互作用组的这一综合分析表明,其在调节少突胶质细胞稳态中的新机制,并为与TET1相关的分子途径提供了全面的见解。我们进一步验证了与选定合作伙伴的TET1互动。总体而言,对少突胶质细胞分化过程中内源性TET1相互作用组的这一综合分析表明,其在调节少突胶质细胞稳态中的新机制,并为与TET1相关的分子途径提供了全面的见解。
更新日期:2020-09-16
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