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Probing Metal-Dependent Phosphate Binding for the Catalysis of the 17E DNAzyme
| Description: | The RNA-cleaving 17E DNAzyme exhibits different levels of cleavage activity in the presence of various divalent metal ions, with Pb2+ giving the fastest cleavage. In this study, the metal–phosphate interaction is probed to understand the trend of activity with different metal ions. For the first-row transition metals, the lowest activity shown by Ni2+ correlates with the inhibition by the inorganic phosphate and its water ligand exchange rate, suggesting inner-sphere metal coordination. Cleavage activity with the two stereoisomers of the phosphorothioate-modified substrates, Rp and Sp, indicated that Mg2+, Mn2+, Fe2+, and Co2+ had the highest Sp:Rp activity ratio of >900. Comparatively, the activity was much less affected using the thiophilic metals, including Pb2+, suggesting inner-sphere coordination. The pH–rate profiles showed that Pb2+ was different than the rest of the metal ions in having a smaller slope and a similar fitted apparent pKa and the pKa of metal-bound water. Combining previous reports and our current results, we propose that Pb2+ most likely plays the role of a general acid while the other metal ions are Lewis acid catalysts interacting with the scissile phosphate. |
| Notes: | Dataset is from publication "Probing Metal-Dependent Phosphate Binding for the Catalysis of the 17E DNAzyme" Item exited embargo and became publicly available on 2022-06-10 |
| Authors: | Moon, Woohyun Jennifer; University of Waterloo Huang, Po-Jung Jimmy; University of Waterloo;  0000-0003-3436-9968Liu, Juewen; University of Waterloo |
| Keywords: | DNAzyme Pb2+ Biosensor |
| Field of Research: | Biological sciences > Biochemistry > Analytical biochemistry
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| Publication Date: | 2022-06-10 |
| Publisher: | Federated Research Data Repository / dépôt fédéré de données de recherche |
| Funder: | Natural Sciences and Engineering Research Council of Canada; Canada First Research Excellence Fund |
| URI: | https://doi.org/10.20383/103.0588 |
| Related Identifiers: |
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| Appears in Collections: | Water Institute |
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Creative Commons Public Domain Dedication (CC0 1.0) https://creativecommons.org/publicdomain/zero/1.0/
Citation
Moon, W., Huang, P., Liu, J. (2022). Probing Metal-Dependent Phosphate Binding for the Catalysis of the 17E DNAzyme. Federated Research Data Repository. https://doi.org/10.20383/103.0588
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