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

Chem. J. Chinese Universities ›› 2011, Vol. 32 ›› Issue (11): 2548.

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

ZHANG Xiao-Ping¹, HUANG Yan-Qin^1*, FAN Chun-Hai^1,2, HUANG Wei^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.

Abstract

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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ZHANG Xiao-Ping, HUANG Yan-Qin*, FAN Chun-Hai, HUANG Wei*. Novel Method of Phosphatase Detection Based on Cationic Conjugated Polymer and Enzymatic Substrate Probe[J]. Chem. J. Chinese Universities, 2011, 32(11): 2548.

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

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