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A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))

The selection of modified DNAzymes represents an important endeavor in expanding the chemical and catalytic properties of catalytic nucleic acids. Few examples of such exist and to date, there is no example where three different modified bases have been simultaneously incorporated for catalytic acti...

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Autores principales: Hollenstein, Marcel, Hipolito, Christopher J., Lam, Curtis H., Perrin, David M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2655665/
https://www.ncbi.nlm.nih.gov/pubmed/19153138
http://dx.doi.org/10.1093/nar/gkn1070
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author Hollenstein, Marcel
Hipolito, Christopher J.
Lam, Curtis H.
Perrin, David M.
author_facet Hollenstein, Marcel
Hipolito, Christopher J.
Lam, Curtis H.
Perrin, David M.
author_sort Hollenstein, Marcel
collection PubMed
description The selection of modified DNAzymes represents an important endeavor in expanding the chemical and catalytic properties of catalytic nucleic acids. Few examples of such exist and to date, there is no example where three different modified bases have been simultaneously incorporated for catalytic activity. Herein, dCTP, dATP and dUTP bearing, respectively, a cationic amine, an imidazole and a cationic guanidine, were enzymatically polymerized on a DNA template for the selection of a highly functionalized DNAzyme, called DNAzyme 9-86, that catalyzed (M(2+))-independent self-cleavage under physiological conditions at a single ribo(cytosine)phosphodiester linkage with a rate constant of (0.134 ± 0.026) min(−1). A pH rate profile analysis revealed pK(a)'s of 7.4 and 8.1, consistent with both general acid and base catalysis. The presence of guanidinium cations permits cleavage at significantly higher temperatures than previously observed for DNAzymes with only amines and imidazoles. Qualitatively, DNAzyme 9-86 presents an unprecedented ensemble of synthetic functionalities while quantitatively it expresses one of the highest reported values for any self-cleaving nucleic acid when investigated under M(2+)-free conditions at 37°C.
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spelling pubmed-26556652009-04-01 A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+)) Hollenstein, Marcel Hipolito, Christopher J. Lam, Curtis H. Perrin, David M. Nucleic Acids Res Chemistry and Synthetic Biology The selection of modified DNAzymes represents an important endeavor in expanding the chemical and catalytic properties of catalytic nucleic acids. Few examples of such exist and to date, there is no example where three different modified bases have been simultaneously incorporated for catalytic activity. Herein, dCTP, dATP and dUTP bearing, respectively, a cationic amine, an imidazole and a cationic guanidine, were enzymatically polymerized on a DNA template for the selection of a highly functionalized DNAzyme, called DNAzyme 9-86, that catalyzed (M(2+))-independent self-cleavage under physiological conditions at a single ribo(cytosine)phosphodiester linkage with a rate constant of (0.134 ± 0.026) min(−1). A pH rate profile analysis revealed pK(a)'s of 7.4 and 8.1, consistent with both general acid and base catalysis. The presence of guanidinium cations permits cleavage at significantly higher temperatures than previously observed for DNAzymes with only amines and imidazoles. Qualitatively, DNAzyme 9-86 presents an unprecedented ensemble of synthetic functionalities while quantitatively it expresses one of the highest reported values for any self-cleaving nucleic acid when investigated under M(2+)-free conditions at 37°C. Oxford University Press 2009-04 2009-01-19 /pmc/articles/PMC2655665/ /pubmed/19153138 http://dx.doi.org/10.1093/nar/gkn1070 Text en © 2009 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Chemistry and Synthetic Biology
Hollenstein, Marcel
Hipolito, Christopher J.
Lam, Curtis H.
Perrin, David M.
A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title_full A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title_fullStr A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title_full_unstemmed A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title_short A self-cleaving DNA enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (M(2+))
title_sort self-cleaving dna enzyme modified with amines, guanidines and imidazoles operates independently of divalent metal cations (m(2+))
topic Chemistry and Synthetic Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2655665/
https://www.ncbi.nlm.nih.gov/pubmed/19153138
http://dx.doi.org/10.1093/nar/gkn1070
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