AID associated protein LC-MS .csv

To identify the protein(s) affecting the activity of AID, we tried to identify the AID interacting proteins by expressing AID-FLAG in 293T cells. The AID interacting proteins were subject to co-immunoprecipitation with anti-FLAG-conjugated beads and then separated by SDS-PAGE. Multiple silver-stained bands, especially proteins over 100 kDa were observed.The visualized bands were excised and examined by mass spectrometry after trypsin digestion. In total 1194 proteins were identified. Among the interactors, 17 proteins were AID-specific interacting proteins.We were especially interested in the factors involved in PTMs.

为鉴定可调控激活诱导胞苷脱氨酶（Activation-Induced Cytidine Deaminase，AID）活性的蛋白质，我们通过在293T细胞中表达带FLAG标签的AID（AID-FLAG），以此鉴定与AID互作的蛋白质。采用抗FLAG偶联磁珠对AID互作蛋白进行免疫共沉淀（co-immunoprecipitation）富集，随后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）完成蛋白分离。银染显色后可观察到多条蛋白条带，其中分子量大于100 kDa的蛋白条带尤为显著。将显色可见的蛋白条带切取后，经胰蛋白酶消化，再通过质谱分析法完成鉴定。本次实验共鉴定得到1194个蛋白质，其中17个为AID特异性互作蛋白。我们对参与蛋白质翻译后修饰（Post-translational modifications，PTMs）的调控因子尤为关注。