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Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures

Given the essential role of apurinic/apyrimidinic endonuclease (APE1) in gene repair and cancer progression, we report a novel approach for probing and regulating cellular APE1 activity by using DNA tetrahedrons. The tetrahedron with an AP site-containing antenna exhibits high sensitivity and specif...

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Autores principales: Zhang, Yi, Deng, Yingnan, Wang, Congshan, Li, Lidan, Xu, Lida, Yu, Yingjie, Su, Xin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6566069/
https://www.ncbi.nlm.nih.gov/pubmed/31360402
http://dx.doi.org/10.1039/c9sc01912j
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author Zhang, Yi
Deng, Yingnan
Wang, Congshan
Li, Lidan
Xu, Lida
Yu, Yingjie
Su, Xin
author_facet Zhang, Yi
Deng, Yingnan
Wang, Congshan
Li, Lidan
Xu, Lida
Yu, Yingjie
Su, Xin
author_sort Zhang, Yi
collection PubMed
description Given the essential role of apurinic/apyrimidinic endonuclease (APE1) in gene repair and cancer progression, we report a novel approach for probing and regulating cellular APE1 activity by using DNA tetrahedrons. The tetrahedron with an AP site-containing antenna exhibits high sensitivity and specificity to APE1. It is suitable for APE1 in vitro detection (detection limit 5 pM) and cellular fluorescence imaging without any auxiliary transfection reagents, which discriminates the APE1 expression level of cancer cells and normal cells. In contrast, the tetrahedron with an AP site on its scaffold exhibits high binding affinity to APE1 but limits enzymatic catalysis making this nanostructure an APE1 inhibitor with an IC(50) of 14.8 nM. It suppresses the APE1 activity in living cells and sensitizes cancer cells to anticancer drugs. We also demonstrate that the APE1 probe and inhibitor can be switched allosterically via stand displacement, which holds potential for reversible inhibition of APE1. Our approach provides a new way for fabricating enzyme probes and regulators and new insights into enzyme–substrate interactions, and it can be expanded to regulate other nucleic acid related enzymes.
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spelling pubmed-65660692019-07-29 Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures Zhang, Yi Deng, Yingnan Wang, Congshan Li, Lidan Xu, Lida Yu, Yingjie Su, Xin Chem Sci Chemistry Given the essential role of apurinic/apyrimidinic endonuclease (APE1) in gene repair and cancer progression, we report a novel approach for probing and regulating cellular APE1 activity by using DNA tetrahedrons. The tetrahedron with an AP site-containing antenna exhibits high sensitivity and specificity to APE1. It is suitable for APE1 in vitro detection (detection limit 5 pM) and cellular fluorescence imaging without any auxiliary transfection reagents, which discriminates the APE1 expression level of cancer cells and normal cells. In contrast, the tetrahedron with an AP site on its scaffold exhibits high binding affinity to APE1 but limits enzymatic catalysis making this nanostructure an APE1 inhibitor with an IC(50) of 14.8 nM. It suppresses the APE1 activity in living cells and sensitizes cancer cells to anticancer drugs. We also demonstrate that the APE1 probe and inhibitor can be switched allosterically via stand displacement, which holds potential for reversible inhibition of APE1. Our approach provides a new way for fabricating enzyme probes and regulators and new insights into enzyme–substrate interactions, and it can be expanded to regulate other nucleic acid related enzymes. Royal Society of Chemistry 2019-05-06 /pmc/articles/PMC6566069/ /pubmed/31360402 http://dx.doi.org/10.1039/c9sc01912j Text en This journal is © The Royal Society of Chemistry 2019 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0)
spellingShingle Chemistry
Zhang, Yi
Deng, Yingnan
Wang, Congshan
Li, Lidan
Xu, Lida
Yu, Yingjie
Su, Xin
Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title_full Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title_fullStr Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title_full_unstemmed Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title_short Probing and regulating the activity of cellular enzymes by using DNA tetrahedron nanostructures
title_sort probing and regulating the activity of cellular enzymes by using dna tetrahedron nanostructures
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6566069/
https://www.ncbi.nlm.nih.gov/pubmed/31360402
http://dx.doi.org/10.1039/c9sc01912j
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