Abstract
Mesoporphyrin IX (MPIX) contains a planar macrocycle center that can interact with various divalent metal ions through the exposed binding sites, leading to the metalation of MPIX. The DNA aptamers for porphyrin molecules usually display different catalytic functions (termed deoxyribozymes or DNAzymes), which can accelerate such chemical reactions. Inspired by this, an affinity chromatography selection approach was designed for identifying a porphyrin metalation DNAzyme. In our experiment, N-methyl mesoporphyrin IX (NMM), an analog of MPIX, is used as the target molecule, owing to its stable and high fluorescence enhancement after combining with specific oligonucleotides. Our results showed that the selected aptamer Nm1 is capable of binding to NMM with a low micromolar dissociation constant (0.75 ± 0.08 μM) and displays a catalytic activity for MPIX metalation with 3.3-fold rate enhancement. The protocol for isolation of such a porphyrin metalation DNAzyme is described in detail here.
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Acknowledgments
This work was financially supported by the Natural Science Foundation of China (21575154, 21775160, 81801837, 31800685), the CAS/SAFEA International Innovation Teams program, and the Science Foundation of Jiangsu Province (BE2016680, BE2018665, BK20180250, BK20180258, BK20180261).
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Yang, L., Cao, Y., Pei, R. (2022). Selection of Aptamer for N-Methyl Mesoporphyrin IX to Develop Porphyrin Metalation DNAzyme. In: Steger, G., Rosenbach, H., Span, I. (eds) DNAzymes. Methods in Molecular Biology, vol 2439. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2047-2_2
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DOI: https://doi.org/10.1007/978-1-0716-2047-2_2
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