高等学校化学学报

高等学校化学学报

• 研究快报 • 上一篇    

Aerolysin纳米孔道探究硫酸乙酰肝素与金属离子的单分子相互作用

张鹏玲1#，高凡2#，陈嘉乐6，邹爱华1，汤娟3，马慧4，蒋翠玲5，万永菁5，夏冰清6，李铁海6，高召兵6，应佚伦2，龙亿涛2
  

  1. 1. 上海师范大学化学与材料科学学院

    2. 南京大学化学学院，分子传感与成像中心，

    3. 江西师范大学化学与材料学院

    4. 浙江理工大学化学与化工学院

    5. 华东理工大学信息科学与工程学院 6. 中国科学院上海药物研究所

  • 收稿日期:2025-12-29 修回日期:2026-01-08 网络首发:2026-01-10 发布日期:2026-01-10
  • 通讯作者: 龙亿涛
  • 基金资助:

    上海市市级科技重大专项和国家自然科学基金委(32250019)资助


Single-Molecule Investigation of Heparan Sulfate-Metal Ion Interactions Using Aerolysin Nanopore

ZHANG Pengling1#, GAO Fan2#, CHEN Jiale6, ZOU Aihua1*, TANG Juan3*, MA Hui4*, JIANG Cuiling5*, WAN Yongjing5*, XIA Bingqing6, LI Tiehai6*, GAO Zhaobing6, YING Yilun2*, LONG Yitao2   

  1. 1. College of Chemistry and Materials Science, Shanghai Normal University

    2. School of Chemistry, Molecular Sensing and Imaging Center, Nanjing University

    3. College of Chemistry and Materials, Jiangxi Normal University

    4. School of Chemistry and Chemical Engineering; Zhejiang Sci-Tech University

    5. School of Information Science and Engineering, East China University of Science and Technology 6. Shanghai Institute of Materia Medica, Chinese Academy of Sciences

  • Received:2025-12-29 Revised:2026-01-08 Online First:2026-01-10 Published:2026-01-10
  • Contact: Yi-Tao Long
  • Supported by:
    Supported by the Shanghai Municipal Science and Technology Major Project, and National Natural Science Foundation of China (No. 32250019)

PDF (PC)

摘要： 肝素(HP)和硫酸乙酰肝素(HS)是一类高度负电性聚糖，通过金属离子介导的蛋白相互作用在多种生物过程中发挥着重要功能。然而，在溶液环境下对HS与金属离子的相互作用进行单分子尺度的直接表征仍极具挑战。本研究利用具有增强静电限域环境的T232K/K238Q Aerolysin突变型纳米孔道，系统研究了HS与不同金属离子之间的相互作用。通过调控电解质中阳离子的种类(Na+、K+和 Ca2+)，揭示了不同金属离子通过调控HS的构象、电荷屏蔽效应以及其与孔道之间的弱相互作用，从而显著影响HS的过孔行为及信号特征。值得注意的是，除了与HS具有较强结合能力的Ca2+外，理化性质相近且结合能力较弱的一价金属离子Na+和K+同样能够诱导HS与孔道作用产生明显差异的单分子信号，展示了纳米孔道技术在单分子尺度解析HP/HS精细结构及其离子依赖性行为方面的独特优势，为HP/HS-金属离子-蛋白质的相互作用解析提供了新的单分子工具。

关键词: 气单胞菌溶素, 单分子分析, 纳米孔道, 硫酸乙酰肝素, 金属离子

Abstract: Heparin (HP) and heparan sulfate (HS) are highly anionic glycosaminoglycans that play essential roles in diverse biological processes through the metal ion-mediated interactions with proteins. However, direct characterization of HS-metal ion interactions at the single-molecule level in solution remains challenging. Nanopore electrochemistry is a label-free and single-molecule technique that enables direct analysis of individual molecular interaction. In this study, a T232K/K238Q aerolysin nanopore featuring an enhanced electrostatic environment was utilized to investigate the interactions between HS and different metal ions. By systematically varying the electrolyte cations (Na+, K+, and Ca2+), we have found that the metal ions significantly regulate HS translocation behavior by modulating its conformation, charge screening, and HS-nanopore interactions. Notably, in addition to Ca2+, which exhibits strong binding affinity to HS, the monovalent cations Na+ and K+ with similar physicochemical properties and weaker binding also induce distinct single-molecule signal signatures. These findings highlight the key role of metal ions in regulating HS-protein interactions and establish the aerolysin nanopore as a sensitive single-molecule platform for resolving ion-dependent interactions of HS in solution.

Key words: Aerolysin, Single-molecule sensing, Nanopore, Heparan sulfate, Metal ion

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