Monitoring DNA–Ligand Interactions in Living Human Cells Using NMR Spectroscopy
| Autor: | Michaela Krafčíková, Silvie Foldynová-Trantírková, Coralie Caron, Tomáš Loja, Tomas Fessl, Radovan Fiala, Simon Dzatko, Robert Hänsel-Hertsch, Jean-Louis Mergny, Marie-Paule Teulade-Fichou, Anton Granzhan, Lukáš Trantírek |
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| Přispěvatelé: | Chimie, Modélisation et Imagerie pour la Biologie [Orsay], Université Paris-Sud - Paris 11 (UP11)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Européen de Chimie et Biologie (IECB), Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cell Survival
Naphthalenes Ligands 010402 general chemistry 01 natural sciences Biochemistry Catalysis Cell Line chemistry.chemical_compound Colloid and Surface Chemistry Anti-Infective Agents Drug Discovery Humans [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Base Pairing Nuclear Magnetic Resonance Biomolecular Binding Sites Ligand Netropsin Biological activity DNA General Chemistry Nuclear magnetic resonance spectroscopy 0104 chemical sciences [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry genomic DNA chemistry Nucleic acid Biophysics Nucleic Acid Conformation Intracellular |
| Zdroj: | Journal of the American Chemical Society Journal of the American Chemical Society, American Chemical Society, 2019, 141 (34), pp.13281-13285. ⟨10.1021/jacs.9b03031⟩ |
| ISSN: | 0002-7863 1520-5126 |
| Popis: | Studies on DNA-ligand interactions in the cellular environment are problematic due to the lack of suitable biophysical tools. To address this need, we developed an in-cell NMR-based approach for monitoring DNA-ligand interactions inside the nuclei of living human cells. Our method relies on the acquisition of NMR data from cells electroporated with preformed DNA-ligand complexes. The impact of the intracellular environment on the integrity of the complexes is assessed based on in-cell NMR signals from unbound and ligand-bound forms of a given DNA target. This technique was tested on complexes of two model DNA fragments and four ligands, namely, a representative DNA minor-groove binder (netropsin) and ligands binding DNA base-pairing defects (naphthalenophanes). In the latter case, we demonstrate that two of the three in vitro-validated ligands retain their ability to form stable interactions with their model target DNA in cellulo, whereas the third one loses this ability due to off-target interactions with genomic DNA and cellular metabolites. Collectively, our data suggest that direct evaluation of the behavior of drug-like molecules in the intracellular environment provides important insights into the development of DNA-binding ligands with desirable biological activity and minimal side effects resulting from off-target binding. |
| Databáze: | OpenAIRE |
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