聚乙烯亚胺修饰的铁酸镍-氧化石墨烯复合物在磷酸化多肽富集中的应用  

Preparation of Polyethylenimine Modified Nickel Ferrite-Graphene Oxide Material and Its Application in Enrichment of Phosphopeptides

作　　者：张恺娜 刘媛 王香凤 刘海灵 谢孟峡 ZHANG Kai-Na;LIU Yuan;WANG Xiang-Feng;LIU Hai-Ling;XIE Meng-Xia(Analytical and Testing Center,Beijing Normal University,Beijing 100875,China;State Key Laboratory of Plant Environmental Resilience,College of Biological Sciences,China Agricultural University,Beijing 100193,China)

机构地区：[1]北京师范大学分析测试中心,北京100875 [2]中国农业大学生物学院,植物抗逆高效全国重点实验室,北京100193

出　　处：《分析化学》2024年第5期664-673,共10页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.22276014,21976016)资助。

摘　　要：制备了一种聚乙烯亚胺(PEI)修饰的铁酸镍-氧化石墨烯复合物材料GO@PEI-NiFe_(2)O_(4),用于选择性净化和富集生物样品中的磷酸化肽段。此复合材料表面的Ni^(2+)和Fe^(3+)可与磷酸基团配位,从而选择性地吸附磷酸化肽段,修饰的PEI增强了复合材料的亲水性,有利于其与磷酸化肽段结合。此外,复合材料中的NiFe_(2)O_(4)具有磁性,可使其快速从溶液中分离出来。以β-酪蛋白(β-Casein)为样品,考察了复合材料对胰酶酶解液中的磷酸化肽段的富集性能,并与GO@NiFe_(2)O_(4)(未被PEI修饰)富集的结果进行了比较,同时探讨了复合材料的吸附机理。采用磷酸化肽段pTyr测定了复合材料对pTyr的静态和动态吸附曲线,最大吸附容量可达36.2μg/mg。研究结果表明,此复合材料能够有效去除非磷酸化肽段等干扰组分的影响,高效地富集复杂基体中的磷酸化肽段。大鼠肝脏蛋白胰酶酶解液经过富集后,采用质谱可鉴定出1535条磷酸化肽段,其富集效果显著优于Fe^(3+)-IMAC商品化试剂盒。本研究为磷酸化肽段的富集提供了一种高选择性材料,在磷酸化蛋白质组学研究中具有潜在的应用前景。The NiFe_(2)O_(4)-graphene oxide nanocomposite modified with polyethylenimine(GO@PEI-NiFe_(2)O_(4))was prepared to purify and enrich phosphopeptides from biosamples.The Ni^(2+)and Fe^(3+)ions on its surface could coordinate with phosphate groups and then selectively adsorb phosphopeptides.PEI was conducive to the above combination due to its high hydrophilicity.The material showed good magnetic response properties and could be rapidly separated from samples with the aid of magnet.With tryptic digest ofβ-casein as sample,the enrichment property of the material to phosphopeptides was studied,which was compared with the results of GO@NiFe_(2)O_(4),revealing the adsorption mechanism of GO@PEI-NiFe_(2)O_(4).The static and dynamic binding properties of GO@PEINiFe_(2)O_(4) were investigated using pTyr as a representative phosphopeptide,and the adsorption capacity was 36.2μg/mg.The results showed that the material could remove the interference of nonphosphopeptides and effectively enrich phosphopeptides in complex matrix.After enrichment by GO@PEI-NiFe_(2)O_(4),1535 phosphopeptides were identified from the tryptic digest of rat liver by mass spectrum and the enrichment effect of GO@PEI-NiFe_(2)O_(4) greatly outperformed commercial Fe^(3+)-IMAC kits.This work provided an efficient material for the enrichment of phosphopeptides,showing potential applications in phosphoproteomics research.

关 键 词：磷酸化肽段 铁酸镍 聚乙烯亚胺 氧化石墨烯 磁响应 

分 类 号：O652.6[理学—分析化学] Q503[理学—化学]