突变KRAS的互作化学分子研究

Study on Chemical Interactors of Mutant KRAS

利用互作蛋白质组化学技术发现G12D突变型KRAS相互作用化学分子。建立融合蛋白质表达量相同的BioID-G12D-KRAS与BioID-WTKRAS细胞,通过LC-MS/MS检测生物素化肽段,定量分析蛋白质组学数据,预测野生型KRAS和G12D突变型KRAS的潜在结合分子。最终成功培养BioID-G12D-KRAS和BioID-WT-KRAS表达含量相同的SW48细胞;定量质谱识别BioID-WT-KRAS组中有88个生物素化蛋白质,BioID-G12DKRAS组中有90个生物素化蛋白质;发现AHNAK2与野生型KRAS更稳定结合,SLC5A3与G12D突变型KRAS优势结合。

To globally discovery the molecules interacting with G12D mutant KRAS by the interaction proteomics technology.The cell system of BioID-G12D-KRAS and BioID-WT-KRAS with the same expression level of fusion protein was established.Biotinylated peptides were detected by LC-MS/MS,and then the proteomics data were quantitatively analyzed.Potential binding molecules of wild-type KRAS and G12D mutant KRAS were predicted.SW48 cells expressing BioID-G12D-KRAS and BioID-WT-KRAS separately were successfully cultured,and both cells expressed KRAS fusion protein in equal amounts.Quantitative mass spectrometry identified 88 biotinylated proteins in the BioID-WT-KRAS group.There were 90 biotinylated proteins in the BioID-G12D-KRAS group.It shows that ANHAK2 binds to KRAS more stably,and SLC5A3 tends to bind with G12D mutant KRAS.