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Development of a Novel Fluorescence Assay Based on the Use of the Thrombin-Binding Aptamer for the Detection of O6-Alkylguanine-DNA Alkyltransferase Activity
Joint Authors
Fàbrega, Carme
Tintoré, Maria
Aviñó, Anna
Ruiz, Federico M.
Eritja, Ramon
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-09-14
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
Human O6-alkylguanine-DNA alkyltransferase (hAGT) is a DNA repair protein that reverses the effects of alkylating agents by removing DNA adducts from the O6 position of guanine.
Here, we developed a real-time fluorescence hAGT activity assay that is based on the detection of conformational changes of the thrombin-binding aptamer (TBA).
The quadruplex structure of TBA is disrupted when a central guanine is replaced by an O6-methyl-guanine.
The sequence also contains a fluorophore (fluorescein) and a quencher (dabsyl) attached to the opposite ends.
In the unfolded structure, the fluorophore and the quencher are separated.
When hAGT removes the methyl group from the central guanine of TBA, it folds back immediately into its quadruplex structure.
Consequently, the fluorophore and the quencher come into close proximity, thereby resulting in decreased fluorescence intensity.
Here, we developed a new method to quantify the hAGT without using radioactivity.
This new fluorescence resonance energy transfer assay has been designed to detect the conformational change of TBA that is induced by the removal of the O6-methyl group.
American Psychological Association (APA)
Tintoré, Maria& Aviñó, Anna& Ruiz, Federico M.& Eritja, Ramon& Fàbrega, Carme. 2010. Development of a Novel Fluorescence Assay Based on the Use of the Thrombin-Binding Aptamer for the Detection of O6-Alkylguanine-DNA Alkyltransferase Activity. Journal of Nucleic Acids،Vol. 2010, no. 2010, pp.1-9.
https://search.emarefa.net/detail/BIM-486716
Modern Language Association (MLA)
Tintoré, Maria…[et al.]. Development of a Novel Fluorescence Assay Based on the Use of the Thrombin-Binding Aptamer for the Detection of O6-Alkylguanine-DNA Alkyltransferase Activity. Journal of Nucleic Acids No. 2010 (2010), pp.1-9.
https://search.emarefa.net/detail/BIM-486716
American Medical Association (AMA)
Tintoré, Maria& Aviñó, Anna& Ruiz, Federico M.& Eritja, Ramon& Fàbrega, Carme. Development of a Novel Fluorescence Assay Based on the Use of the Thrombin-Binding Aptamer for the Detection of O6-Alkylguanine-DNA Alkyltransferase Activity. Journal of Nucleic Acids. 2010. Vol. 2010, no. 2010, pp.1-9.
https://search.emarefa.net/detail/BIM-486716
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-486716