大学化学 >> 2021, Vol. 36 >> Issue (12): 2102062.doi: 10.3866/PKU.DXHX202102062

化学实验 上一篇    下一篇

电喷雾多级质谱研究功能小分子与蛋白质的非共价相互作用——推荐一个化学生物学教学实验

张于锰, 陈语嫣, 刘艳()   

  • 收稿日期:2021-02-28 录用日期:2021-03-30 发布日期:2021-04-23
  • 通讯作者: 刘艳 E-mail:stacyliu@xmu.edu.cn
  • 作者简介:刘艳, Email: stacyliu@xmu.edu.cn
  • 基金资助:
    国家人才培养基金项目(J0630429)

A Chemical Biology Teaching Experiment: Characterization of the Non-Covalent Interaction between Functional Small Molecule and Proteins by Electrospray Multistage Mass Spectrometry

Yumeng Zhang, Yuyan Chen, Yan Liu()   

  • Received:2021-02-28 Accepted:2021-03-30 Published:2021-04-23
  • Contact: Yan Liu E-mail:stacyliu@xmu.edu.cn

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摘要：

肌红蛋白不仅便宜易得，而且自身就是一种由蛋白质和铁卟啉环辅基通过非共价相互作用构成的复合物。因此，以肌红蛋白为研究模型，带领学生实践基于电喷雾多级质谱技术的酶抑制剂直接筛选策略，重点考查不同有机溶剂比例及pH等样品制备条件对复合物测定结果的影响，以及如何利用二级质谱技术实现功能小分子配体的在线定性分析。该实验已在我校化学生物学专业本科生“化学生物学综合实验”教学工作中进行了多轮教学实践，通过实践不断优化实验设计，取得了良好的教学效果。

关键词: 电喷雾多级质谱, 非共价相互作用, 肌红蛋白

Abstract:

Myoglobin is not only easily available, but also an inherent complex between protein and porphyrin ligand. Therefore, non-covalent complex of myoglobin was selected as the research model for teaching experiment to lead the senior students to practice the direct screening strategy of enzyme inhibitors by electrospray multistage mass spectrometry. Through comparing the effects of different sample preparation conditions on the MS determination of the complex, including organic solvent content and pH, students will understand the importance of sample preparation for MS analysis and grasp how to use MS/MS technology to achieve online qualitative analysis of functional small molecule ligands. The experiment here has been used for a few years as one of the teaching experiments in "comprehensive chemical biology experiments" course for undergraduates majoring in chemical biology at our college. With continuous teaching practice and optimization of the experiment, some good teaching effects have been achieved.

Key words: Electrospray multistage mass spectrometry, Non-covalent interaction, Myoglobin