Identification of Novel MAGE-G1-Interacting Partners in Retinoic Acid-Induced P19 Neuronal Differentiation Using SILAC-Based Proteomics.
Identification of Novel MAGE-G1-Interacting Partners in Retinoic Acid-Induced P19 Neuronal Differentiation Using SILAC-Based Proteomics.
复制标题
使用基于 SILAC 的蛋白质组学鉴定视黄酸诱导的 P19 神经元分化中的新型 MAGE-G1 相互作用伙伴。
DOI：
10.1038/srep44699
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Liu Shaojun
中科院分区：
文献类型：
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作者：
Liu Yong;Chen Yujian;Lin Shide;Yang Shuguang;Liu Shaojun
MAGE-G1是一种在神经发生的早期过程中发挥作用的蛋白质。然而，MAGE-G1在神经发生中扮演的基本角色还没有完全被理解。寻找与MAGE-G1相互作用的伴侣，将有助于MAGE-G1的功能研究。本研究利用细胞培养-免疫沉淀定量蛋白质组学中的氨基酸稳定同位素标记技术，筛选了视黄酸诱导P19细胞分化过程中MAGE-G1的相互作用蛋白，首次发现FSCN1和VIME是潜在的新型MAGE-G1相互作用蛋白。通过细菌GST-down实验和COS7细胞免疫共沉淀实验验证MAGE-G1与FSCN1或VIME的相互作用。内源性免疫共沉淀实验进一步证实，在维甲酸诱导P19细胞分化6天后，MAGE-G1与FSCN1或VIME相互作用。这些结果提供了MAGE-G1与FSCN1或VIME之间的功能联系，并可能有助于更好地理解MAGE-G1在神经发生过程中的基本方面。
MAGE-G1 is a protein plays role in the early process of neurogenesis. However, the fundamental roles MAGE-G1 played in neurogenesis have not yet been completely understood. Finding the partners MAGE-G1 interacting with will surely contribute to the function study of MAGE-G1. In this study, using Stable Isotope Labeling by Amino acids in Cell culture-immunoprecipitation quantitative proteomics, we screened the interacting proteins of MAGE-G1 during retinoic acid -induced neuronal differentiation of P19 cells and firstly found that FSCN1 and VIME were potential novel MAGE-G1-interacting proteins. Then, the interaction between overexpressed MAGE-G1 and FSCN1 or VIME was validated by GST-pull down assay in bacteria and by co-immunoprecipitation assay in COS7 cells. Endogenous co-immunoprecipitation assay further confirmed that MAGE-G1 interacted with FSCN1 or VIME in P19 cells after a 6-day retinoic acid-induced neuronal differentiation. Those results provide a functional linkage between MAGE-G1 and FSCN1 or VIME and may facilitate a better understanding of the fundamental aspects of MAGE-G1 during neurogenesis.