基于天冬氨酸的固定化金属离子亲和色谱材料用于磷酸化肽选择性富集

摘要

采用点击化学的方法将自然界中的天冬氨酸(aspartic acid)键合到硅球上(命名为 Click Asp),并将 Fe3+配位到 Click Asp上,合成固定金属离子亲和色谱(IMAC)材料(Fe3+-Click Asp);采用红外光谱、X射线电子能谱和扫描电镜等表征证明 Fe3+-Click Asp成功合成.将此 IMAC材料用于蛋白质酶解液和牛奶中的磷酸化肽的富集,实现了磷酸化肽的高选择性富集.本研究为磷酸化蛋白质组学提供了新材料和新方法.%A natural amino acid,aspartic acid,was modified to bind to silica microspheres with a click chemistry method(-Click Asp). The modified Click Asp microspheres were further modified by incorporating Fe3+ions,resulting in Fe3+and Click Asp based immobilized ion affin-ity chromatography(IMAC)(Fe3+-Click Asp). The prepared materials were characterized using Fourier transform infrared spectrometry,X-ray photoelectron spectrometry,and scanning elec-tron microscopy,and it was confirmed that Fe3+-Click Asp was successfully prepared. Subse-quently,the Fe3+-Click Asp microspheres were used to enrich phosphopeptides from tryptic digests of proteins and milk. It was found that the phosphopeptides could be selectively enriched with this material. This study describes a novel material and enrichment method for phosphoproteomics.