Spectrofluorometry of Ions and Small Molecules Based on Conformational Changes of Specific Oligonucleotides with o-Phthalaldehyde-β-mercaptoethanol

Chemical Research in Chinese Universities ›› 2010, Vol. 26 ›› Issue (3): 360-365.

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Spectrofluorometry of Ions and Small Molecules Based on Conformational Changes of Specific Oligonucleotides with o-Phthalaldehyde-β-mercaptoethanol

LIAO Qie-gen^1,2, WANG Jian¹, LONG Yun-fei¹ and LI Yuan-fang^1*

1. Key Laboratory of Luminescence and Real-Time Analysis, Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, P. R. China;
2. Key Laboratory of Quality and Safety of Agricultural Products, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, P. R. China

Received:2009-06-10 Revised:2009-10-15 Online:2010-05-25 Published:2010-07-27
Contact: LI Yuan-fang. E-mail: liyf@swu.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China(No.20775061).

Abstract

Abstract:

Specific oligonucleotides such as telomere DNA and aptamer often undergo conformational changes upon ligand binding. Composite reagent composed of o-phthalaldehyde and β-mercaptoethanol(OPAME) has been extensively applied to fluorescent detection of amino compounds based on the reaction of primary amino-group, herein we proposed a general spectrofluorometry for ions and small molecules due to conformational changes upon ligand binding taking K⁺ and ATP as examples. In a borate controlled buffer medium, telomere DNA could react with OPAME, giving a thio-subtituted isoindole compound with strong fluorescence emission at 455 nm when excited at 340 nm. It was found that however, the fluorescence emission was greatly reduced in the presence of K⁺ since the formation of the quadruplex structure inhibits the reaction activity of amino-groups of telomere DNA. In order to testify the general application of OPAME reagent based on the conformational change of oligonucleotides, we further proposed a sensitive method of ATP based on its highly selective interaction with ATP-aptamer. The above mentioned applications show that the spectrofluorometry with the aid of OPAME reagent is simple, label free that is expected to be potentially general for DNA conformational change-based target detection.

Key words: Aptamer; o-Phthalaldehyde; β-Mercaptoethanol; K+; Adenosine-5′-triphosphoric acid

LIAO Qie-gen, WANG Jian, LONG Yun-fei and LI Yuan-fang*. Spectrofluorometry of Ions and Small Molecules Based on Conformational Changes of Specific Oligonucleotides with o-Phthalaldehyde-β-mercaptoethanol[J]. Chemical Research in Chinese Universities, 2010, 26(3): 360-365.

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Spectrofluorometry of Ions and Small Molecules Based on Conformational Changes of Specific Oligonucleotides with o-Phthalaldehyde-β-mercaptoethanol

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