COP9信号小体（CSN）通过参与不同信号通路影响诸多重要的生物学功能，如细胞周期调控、DNA损伤修复和肿瘤发生等。近年来，CSN因为与IKK复合物的相互作用而被链接到NF-κB信号通路，但是具体的调控机制仍不清楚。我们前期研究发现IKK的过量表达会导致CSN的活性亚基CSN5磷酸化，并通过高精度的质谱分析确定IKK诱导的CSN5的S201和T205磷酸化会损害CSN的deneddylation活性。同时，我们发现TNF-α诱导的IKK活化具有类似效应，并导致广泛的CSN作用蛋白网络的变化。因此，证明IKK可以直接磷酸化CSN5，并阐明这一事件对于CSN活性和作用蛋白网络的影响至关重要。运用基于质谱的QTAX策略和QconCAT技术，监测TNF-α刺激后CSN的亚基组成、翻译后修饰以及重要结合蛋白和蛋白复合物的影响，从而为CSN相关的疾病或肿瘤发生机理提供解释，并为其治疗提供靶点。

The COP9 signalosome (CSN) has been reported to participate in various signaling pathways to regulate diverse biological processes, including cell cycle progression, DNA damage repair and tumorgenesis. Recently, CSN complex was linked to the NF-κB signaling pathway due to its interaction with the IKK complex. Nevertheless, how the CSN complex is regulated during the NF-κB activation remains unclear. In our preliminary study, we determined that overexpression of IKKs induced enhanced phosphorylation of the CSN5 subunit, which harbors the catalytic site of CSN-mediated deneddylation reactions. Through mutational analyses, it appeared that phosphorylation at serine 201 and threonine 205 of CSN5 impairs CSN-mediated deneddylation. Meanwhile, we found that TNF-α induced IKK activation could also cause the phosphorylation of CSN5 subunit, and lead to the extensive interaction network change of CSN. Thus, in our future study, we are expecting to verify that IKK could directly phosphorylate CSN5. By employing mass spectrometry based QTAX strategy and QconCAT technology, we will be able to high-throughput monitor the change of CSN interaction work at different time points post TNF-α stimulation, dynamically analyze the effects of TNF-α on subunits composition, post-translational modifications and crucial associating proteins of CSN, which would provide explanation for the mechanism of CSN related diseases or tumors and targets for their therapy.