Photo-crosslinking proteomic analysis of IFITM3 interactors

A diazirine-containing unnatural amino acid was site-specifically incorporated into IFITM3, an antiviral protein. After photo-irradiation, the interacting proteins were crosslinked with IFITM3, isolated by immunoprecipitation, and identified by quantitative SILAC proteomics.将含双吖丙啶（diazirine）的非天然氨基酸位点特异性整合至干扰素诱导跨膜蛋白3（IFITM3，一种抗病毒蛋白）中。经光辐照处理后，与IFITM3存在相互作用的蛋白可与其发生交联，随后通过免疫沉淀法分离，并采用稳定同位素标记氨基酸细胞培养（SILAC）定量蛋白质组学技术完成鉴定。