Conjugated polyelectrolyte based real-time fluorescence assay for adenylate kinase

Analytical Chemistry
Yan Liu, Kirk S Schanze

Abstract

Addition of adenosine 5'-triphosphate (ATP) to a solution of the anionic conjugated polyelectrolyte PPECO2 and copper(II) ion (Cu2+) recovers the Cu2+-quenched fluorescence of PPECO(2) to a significantly greater extent compared with the addition of adenosine 5'-diphosphate (ADP) or adenosine 5'-monophosphate (AMP) at the same concentration levels. Taking advantage of the differential response of the PPECO2-Cu2+ system to ATP, ADP and AMP, we have developed fluorescence turn-off and turn-on assays that monitor the catalytic activity of adenylate kinase (ADK) in the equilibrium transphosphorylation reaction (ATP + AMP <--> 2ADP). The fluorescence turn-on and turn-off assays monitor the forward and reverse transphosphorylation reactions, respectively. The forward assay operates with ATP substrate present at the submillimolar concentration range and offers a straightforward and rapid detection of ADK catalytic activity with the enzyme present in the nanomolar range, in either end-point or real-time formats. The real-time fluorescence intensity from PPECO2 can be converted to substrate (ATP) concentration in the forward reaction assay by using an ex-situ calibration curve, allowing ADK catalyzed reaction rates and kinetic parameters...Continue Reading

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Citations

Sep 5, 2012·Analytical and Bioanalytical Chemistry·Umit Hakan YildizBo Liedberg
Jun 8, 2012·Chemical Reviews·Chunlei ZhuShu Wang
Mar 17, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Yanli TangDavid G Whitten
Jul 27, 2010·Chemical Communications : Chem Comm·Xiaoyong Zhao, Kirk S Schanze
Dec 25, 2015·Chemistry, an Asian Journal·Jin-Zhu GeJiang-Hua Zhu
May 14, 2011·Macromolecular Rapid Communications·Fude FengShu Wang
Oct 27, 2010·Physical Chemistry Chemical Physics : PCCP·Mijeong KangHan Young Woo
May 16, 2009·Angewandte Chemie·Hui JiangKirk S Schanze
Aug 28, 2013·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Atul Kumar Dwivedi, Parameswar Krishnan Iyer
Jul 15, 2009·Chemistry, an Asian Journal·Lingling An, Shu Wang
Mar 14, 2013·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Renxuan LiuChunyan Tan
Jun 17, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Xuli FengShu Wang
Dec 5, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Fang PuXiaogang Qu
Mar 6, 2015·ACS Applied Materials & Interfaces·Yi WuChunyan Tan
May 17, 2012·The Journal of Physical Chemistry Letters·Eric H HillDavid G Whitten
Jul 10, 2013·Langmuir : the ACS Journal of Surfaces and Colloids·Eric H HillDavid G Whitten

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