高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (1): 48-53.doi: 10.7503/cjcu20170312

• 分析化学 • 上一篇    下一篇

重组人促红细胞生成素 DNA适配体的活性高级结构特征

贺小琴, 何军林, 徐华, 郭磊(), 谢剑炜   

  1. 抗毒药物与毒理学国家重点实验室, 军事医学科学院毒物药物研究所, 北京 100850
  • 收稿日期:2017-05-18 出版日期:2018-01-10 发布日期:2017-11-13
  • 作者简介:联系人简介: 郭 磊, 女, 博士, 研究员, 主要从事生物分析化学和毒物药物分析研究. E-mail:guolei@bmi.ac.cn
  • 基金资助:
    国家自然科学基金(批准号: 21175152)资助

Active Conformation of DNA Aptamer Against Recombinant Human Erythropoietin-α

HE Xiaoqin, HE Junlin, XU Hua, GUO Lei*(), XIE Jianwei   

  1. Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, State Key Laboratory of Toxicology and Medical Countermeasures, Beijing 100850, China
  • Received:2017-05-18 Online:2018-01-10 Published:2017-11-13
  • Contact: GUO Lei E-mail:guolei@bmi.ac.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(No.21175152)

摘要:

针对重组人促红细胞生成素-α(EPO-α)的最短适配体序列In27, 使用表面等离子体共振、 圆二色光谱及荧光光谱等多种技术, 研究和明确了In27的活性与构型间的关系. 结果表明, 阳离子Na+的存在及其浓度是影响In27产生特异性亲和识别的关键因素; 在Na+诱导下In27形成了平行/反平行的混合G四链体(GQ)构型; 荧光探针硫磺素T(ThT)能特异性结合该混合构型, EPO-α的加入可与ThT形成竞争, 从而便于利用ThT创建活性EPO-α的“光开关”型检测探针.

关键词: 表面等离子体共振, 圆二色光谱, 荧光探针, G-四链体, Na+

Abstract:

A series of spectroscopic techniques such as surface plasmon resonance, circular dichroism and fluorescence was employed to illustrate the relationship between the binding activity and conformation of the shortest aptamer In27 against recombinant human erythropoietin-α(EPO-α). It was found that the presence and concentration of sodium cation were the key factors that affected the specific affinity recognition of aptamer In27. Sodium cation induced aptamer In27 to form a parallel/antiparallel hybrid G-quadruplex(GQ) conformation. GQ-specific fluorescent probe thysmenonine T(ThT) specifically bound to the hybrid GQ conformation of aptamer In27. The addition of EPO-α could compete with ThT to facilitate to create a label-free fluorescent “turn-off” probe for EPO-α. This research provides an effective combination of techniques for elucidating the active conformation of novel aptamers, and provides a kind of aptameric modules with clear interaction features for analytical, sensing and diagnostic applications.

Key words: Surface plasmon resonance, Circular dichroism, Fluorescent probes, G-Quadruplex, Sodium cation

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