高等学校化学学报

高等学校化学学报 ›› 2011, Vol. 32 ›› Issue (11): 2548.

• 研究论文 • 上一篇    下一篇

基于阳离子型共轭聚合物与酶底物探针的磷酸酯酶检测新方法

张小平1,  黄艳琴1,  樊春海1,2,  黄维1   

  1. 1. 有机电子与信息显示国家重点实验室培育基地, 南京邮电大学信息材料与纳米技术研究院,  南京 210046; 
    2. 中国科学院上海应用物理研究所, 上海 201800
  • 收稿日期:2010-12-28 修回日期:2011-03-16 出版日期:2011-11-10 发布日期:2011-10-14
  • 通讯作者: 黄维;黄艳琴 E-mail:wei-huang@njupt.edu.cn;iamyqhuang@njupt.edu.cn
  • 基金资助:

    国家“九七三”计划项目(批准号: 2009CB930601)、国家自然科学基金(批准号: 20804018, 51073078)、江苏省自然科学基金(批准号: BK2008452)、江苏省高等学校优秀科技创新团队项目(批准号: TJ207035)和江苏省高等学校自然科学研究项目(批准号: 10KJB150014)资助.

Novel Method of Phosphatase Detection Based on Cationic Conjugated Polymer and Enzymatic Substrate Probe

ZHANG Xiao-Ping1, HUANG Yan-Qin1*, FAN Chun-Hai1,2, HUANG Wei1*   

  1. 1.  Key Laboratory for Organic Electronics & Information Displays,  Institute of Advanced Materials, Nanjing University of Posts & Telecommunications, Nanjing 210046, China;
    2.  Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China
  • Received:2010-12-28 Revised:2011-03-16 Online:2011-11-10 Published:2011-10-14
  • Contact: HUANG Wei;HUANG Yan-Qin E-mail:wei-huang@njupt.edu.cn;iamyqhuang@njupt.edu.cn
  • Supported by:

    国家“九七三”计划项目(批准号: 2009CB930601)、国家自然科学基金(批准号: 20804018, 51073078)、江苏省自然科学基金(批准号: BK2008452)、江苏省高等学校优秀科技创新团队项目(批准号: TJ207035)和江苏省高等学校自然科学研究项目(批准号: 10KJB150014)资助.

PDF (PC)

被引次数

9

摘要： 建立了一种基于阳离子型共轭聚合物与酶底物探针的磷酸酯酶检测新方法.  阳离子型共轭聚合物通过静电吸引与荧光素修饰的带负电荷的三磷酸腺苷结合,  并发生荧光能量共振转移; 当加入磷酸酯酶时, 它可以催化底物三磷酸腺苷上的磷酸酯基团逐渐水解, 得到不带电荷的腺苷, 从而使阳离子型共轭聚合物得以释放, 能量转移效率下降. 实验结果表明, 能量转移效率下降的程度与酶的浓度相关. 该方法操作简单, 响应速度快, 灵敏度高, 易于扩展至其它能催化底物电荷密度变化的酶的检测, 而且还有望应用于对酶具有抑制作用的药物分子的筛选.

关键词: 水溶性共轭聚合物, 底物, 磷酸酯酶, 荧光检测, 荧光能量共振转移

Abstract: A novel method was  developed for phosphatase detection based on  cationic conjugated polymer(CCP) and enzymatic substrate.  The CCP associated with the fluorescein-labeled anionic adenosine triphosphate(ATP) by electrostatic attractions and fluorescence resonance energy transfer(FRET) from the CCP to the fluorescein-labeled ATP occurred.  When phosphatase was added, it catalyzed the hydrolysis of phosphate groups from its substrate, ATP, and the final product adenosine was neutral, which made the CCP detached from adenosine.  Then the FRET efficiency decreased.  The results show that the degree of FRET efficiency decrease is related to the concentration of phosphatase.  This detection method has the advantages including easy operations, real-time response, and high sensitivity.  It can also sensitively assay other enzymes that promote the changes of the charge density of substrates; moreover, it provides a promising application in drug screening based on the inhibition of enzymatic activity.

Key words: Water-soluble conjugated polymer, Substrate, Phosphatase, Fluorescence detection, Fluorescence resonance energy transfer

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