纳米粒子表面蛋白构象的交联质谱原位蛋白质组表征  

In-situ Proteomic Characterization of Protein Conformation on Nanoparticle Surface by Cross-linking Mass Spectrometry

作　　者：张红燕 周烨 吴仁安[1] ZHANG Hong-Yan;ZHOU Ye;WU Ren-An(Key Laboratory of Separation Science for Analytical Chemistry,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院大连化学物理研究所,中国科学院分离分析化学重点实验室,大连116023 [2]中国科学院大学,北京100049

出　　处：《分析化学》2018年第10期1570-1580,共11页Chinese Journal of Analytical Chemistry

基　　金：本文系国家自然科学基金项目(Nos.21375125,21675156)资助。

摘　　要：采用基于化学交联质谱(Chemical cross-linking mass spectrometry,CXMS)的蛋白质组学方法研究了四氧化三铁(Fe3O4)纳米粒子表面牛血清白蛋白(BSA)的构象变化,考察了纳米粒子的曲率半径和蛋白孵育浓度与Fe_3O_4纳米粒子表面BSA构象的关系。CXMS结合傅里叶变换衰减全反射红外光谱(Attenuated total reflectance Fourier transform infrared spectroscopy,ATR-FTIR)的蛋白质二级结构信息表明,BSA在Fe_3O_4纳米粒子表面存在构象变化和排列取向现象,反映在BSA接触材料后一些位点交联频率的改变。此纳米粒子表面蛋白的CXMS表征方法简单、快速,可提供蛋白在纳米材料上结构变化的化学交联的束缚特征,有望进一步应用于复杂蛋白体系与材料的相互作用研究。The conformation change of bovine serum albumin(BSA)absorbed on Fe 3O 4 nanoparticle surface was characterized by chemical cross-linking mass spectrometry(CXMS)-based proteomics approach.The effects of nanoparticle surface curvatures and initial protein concentrations on protein conformation change were investigated by the CXMS.The results revealed that the BSA adsorbed on the surface of Fe3O4 nanoparticle could undergo an altered conformational state in comparison to that of BSA in solution as the crosslinking frequency of some amino acid residues of the BSA varied significantly,with the assistant analysis of the attenuated total reflectance Fourier transfer infrared spectroscopy(ATR-FTIR).By virtue of the high-throughput,high accuracy and high sensitivity of mass spectrometry,the CXMS-based proteomic method could provide structural information of proteins adsorbed on nanoparticle surface by crosslinking restraints and would be promised to be applied to the analysis of nano-bio interaction for proteins in complex biological systems.

关 键 词：交联质谱 蛋白质组 纳米材料 蛋白质构象 

分 类 号：O629.73[理学—有机化学] O657.63[理学—化学]