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Characterization of clinically used oral antiseptics as quadruplex-binding ligands

Approaches to characterize the nucleic acid-binding properties of drugs and druglike small molecules are crucial to understanding the behavior of these compounds in cellular systems. Here, we use a Small Molecule Microarray (SMM) profiling approach to identify the preferential interaction between ch...

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Autores principales: Calabrese, David R, Zlotkowski, Katherine, Alden, Stephanie, Hewitt, William M, Connelly, Colleen M, Wilson, Robert M, Gaikwad, Snehal, Chen, Lu, Guha, Rajarshi, Thomas, Craig J, Mock, Beverly A, Schneekloth, John S
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5888870/
https://www.ncbi.nlm.nih.gov/pubmed/29481610
http://dx.doi.org/10.1093/nar/gky084
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author Calabrese, David R
Zlotkowski, Katherine
Alden, Stephanie
Hewitt, William M
Connelly, Colleen M
Wilson, Robert M
Gaikwad, Snehal
Chen, Lu
Guha, Rajarshi
Thomas, Craig J
Mock, Beverly A
Schneekloth, John S
author_facet Calabrese, David R
Zlotkowski, Katherine
Alden, Stephanie
Hewitt, William M
Connelly, Colleen M
Wilson, Robert M
Gaikwad, Snehal
Chen, Lu
Guha, Rajarshi
Thomas, Craig J
Mock, Beverly A
Schneekloth, John S
author_sort Calabrese, David R
collection PubMed
description Approaches to characterize the nucleic acid-binding properties of drugs and druglike small molecules are crucial to understanding the behavior of these compounds in cellular systems. Here, we use a Small Molecule Microarray (SMM) profiling approach to identify the preferential interaction between chlorhexidine, a widely used oral antiseptic, and the G-quadruplex (G4) structure in the KRAS oncogene promoter. The interaction of chlorhexidine and related drugs to the KRAS G4 is evaluated using multiple biophysical methods, including thermal melt, fluorescence titration and surface plasmon resonance (SPR) assays. Chlorhexidine has a specific low micromolar binding interaction with the G4, while related drugs have weaker and/or less specific interactions. Through NMR experiments and docking studies, we propose a plausible binding mode driven by both aromatic stacking and groove binding interactions. Additionally, cancer cell lines harbouring oncogenic mutations in the KRAS gene exhibit increased sensitivity to chlorhexidine. Treatment of breast cancer cells with chlorhexidine decreases KRAS protein levels, while a KRAS gene transiently expressed by a promoter lacking a G4 is not affected. This work confirms that known ligands bind broadly to G4 structures, while other drugs and druglike compounds can have more selective interactions that may be biologically relevant.
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spelling pubmed-58888702018-04-11 Characterization of clinically used oral antiseptics as quadruplex-binding ligands Calabrese, David R Zlotkowski, Katherine Alden, Stephanie Hewitt, William M Connelly, Colleen M Wilson, Robert M Gaikwad, Snehal Chen, Lu Guha, Rajarshi Thomas, Craig J Mock, Beverly A Schneekloth, John S Nucleic Acids Res Chemical Biology and Nucleic Acid Chemistry Approaches to characterize the nucleic acid-binding properties of drugs and druglike small molecules are crucial to understanding the behavior of these compounds in cellular systems. Here, we use a Small Molecule Microarray (SMM) profiling approach to identify the preferential interaction between chlorhexidine, a widely used oral antiseptic, and the G-quadruplex (G4) structure in the KRAS oncogene promoter. The interaction of chlorhexidine and related drugs to the KRAS G4 is evaluated using multiple biophysical methods, including thermal melt, fluorescence titration and surface plasmon resonance (SPR) assays. Chlorhexidine has a specific low micromolar binding interaction with the G4, while related drugs have weaker and/or less specific interactions. Through NMR experiments and docking studies, we propose a plausible binding mode driven by both aromatic stacking and groove binding interactions. Additionally, cancer cell lines harbouring oncogenic mutations in the KRAS gene exhibit increased sensitivity to chlorhexidine. Treatment of breast cancer cells with chlorhexidine decreases KRAS protein levels, while a KRAS gene transiently expressed by a promoter lacking a G4 is not affected. This work confirms that known ligands bind broadly to G4 structures, while other drugs and druglike compounds can have more selective interactions that may be biologically relevant. Oxford University Press 2018-04-06 2018-02-22 /pmc/articles/PMC5888870/ /pubmed/29481610 http://dx.doi.org/10.1093/nar/gky084 Text en Published by Oxford University Press on behalf of Nucleic Acids Research 2018. This work is written by (a) US Government employee(s) and is in the public domain in the US.
spellingShingle Chemical Biology and Nucleic Acid Chemistry
Calabrese, David R
Zlotkowski, Katherine
Alden, Stephanie
Hewitt, William M
Connelly, Colleen M
Wilson, Robert M
Gaikwad, Snehal
Chen, Lu
Guha, Rajarshi
Thomas, Craig J
Mock, Beverly A
Schneekloth, John S
Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title_full Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title_fullStr Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title_full_unstemmed Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title_short Characterization of clinically used oral antiseptics as quadruplex-binding ligands
title_sort characterization of clinically used oral antiseptics as quadruplex-binding ligands
topic Chemical Biology and Nucleic Acid Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5888870/
https://www.ncbi.nlm.nih.gov/pubmed/29481610
http://dx.doi.org/10.1093/nar/gky084
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