Rationally designed aptamer-based fluorescence polarization sensor dedicated to the small target analysis
| Autor: | Valérie Guieu, Sandrine Perrier, Béatrice Roy, Eric Peyrin, Corinne Ravelet, Christian Périgaud, Jennifer Fize |
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| Přispěvatelé: | Département de pharmacochimie moléculaire (DPM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT-FR 2599), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Département de pharmacochimie moléculaire |
| Rok vydání: | 2010 |
| Předmět: |
Adenosine monophosphate
Adenosine Stereochemistry Aptamer Biomedical Engineering Biophysics Biosensing Techniques 010402 general chemistry Sensitivity and Specificity 01 natural sciences chemistry.chemical_compound Electrochemistry [CHIM]Chemical Sciences Fluorescein Protein secondary structure ComputingMilieux_MISCELLANEOUS 010401 analytical chemistry Reproducibility of Results Equipment Design General Medicine Aptamers Nucleotide Small molecule 0104 chemical sciences Equipment Failure Analysis Refractometry Spectrometry Fluorescence chemistry Nucleic acid DNA Probes Biosensor Fluorescence anisotropy Biotechnology |
| Zdroj: | Biosensors and Bioelectronics Biosensors and Bioelectronics, Elsevier, 2010, 25, pp.1652-1657. ⟨10.1016/j.bios.2009.12.005⟩ |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2009.12.005 |
| Popis: | A direct fluorescence polarization (FP) assay strategy, dedicated to the small molecule sensing and based on the unique induced-fit binding mechanism of end-labelled nucleic acid aptamers, has been recently developed by our group. Small target binding has been successfully converted into a significant increase of the fluorescence anisotropy signal presumably produced by the reduction of the local motional freedom of the dye. In order to generalize the approach, a rational FP sensor methodology was established herein, by engineering instability in the secondary structure of an aptameric recognition element. The anti-adenosine DNA aptamer, labelled by a single fluorescein dye at its 3' extremity, was employed as a model functional nucleic acid probe. The terminal stem of the stem-loop structure was shortened to induce a destabilized/denatured conformation which promoted the local segmental mobility of the dye and then a significant depolarization process. Upon target binding, the structural change of the aptamer induced the formation of a stable stem-loop structure, leading to the reduction of the dye mobility and the increase in the fluorescence anisotropy signal. This reasoned approach was applied to the sensing of adenosine and adenosine monophosphate and their chiral analysis. |
| Databáze: | OpenAIRE |
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