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Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs
[Image: see text] Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO(•)) metabolism, storage, and transport as well as mediators in numerous NO-signaling pathways. RSNO levels are finely regulated, and dysregulation is associated with the etiology of several path...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2013
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663071/ https://www.ncbi.nlm.nih.gov/pubmed/23614769 http://dx.doi.org/10.1021/ja401565w |
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author | Seneviratne, Uthpala Godoy, Luiz C. Wishnok, John S. Wogan, Gerald N. Tannenbaum, Steven R. |
author_facet | Seneviratne, Uthpala Godoy, Luiz C. Wishnok, John S. Wogan, Gerald N. Tannenbaum, Steven R. |
author_sort | Seneviratne, Uthpala |
collection | PubMed |
description | [Image: see text] Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO(•)) metabolism, storage, and transport as well as mediators in numerous NO-signaling pathways. RSNO levels are finely regulated, and dysregulation is associated with the etiology of several pathologies. Current methods for RSNO quantification depend on indirect assays that limit their overall specificity and reliability. Recent developments of phosphine-based chemical probes constitute a promising approach for the direct detection of RSNOs. We report here results from a detailed mechanistic and kinetic study for trapping RSNOs by three distinct phosphine probes, including structural identification of novel intermediates and stability studies under physiological conditions. We further show that a triarylphosphine-thiophenyl ester can be used in the absolute quantification of endogenous GSNO in several cancer cell lines, while retaining the elements of the SNO functional group, using an LC–MS-based assay. Finally, we demonstrate that a common product ion (m/z = 309.0), derived from phosphine–RSNO adducts, can be used for the detection of other low-molecular weight nitrosothiols (LMW-RSNOs) in biological samples. Collectively, these findings establish a platform for the phosphine ligation-based, specific and direct detection of RSNOs in biological samples, a powerful tool for expanding the knowledge of the biology and chemistry of NO(•)-mediated phenomena. |
format | Online Article Text |
id | pubmed-3663071 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-36630712013-05-28 Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs Seneviratne, Uthpala Godoy, Luiz C. Wishnok, John S. Wogan, Gerald N. Tannenbaum, Steven R. J Am Chem Soc [Image: see text] Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO(•)) metabolism, storage, and transport as well as mediators in numerous NO-signaling pathways. RSNO levels are finely regulated, and dysregulation is associated with the etiology of several pathologies. Current methods for RSNO quantification depend on indirect assays that limit their overall specificity and reliability. Recent developments of phosphine-based chemical probes constitute a promising approach for the direct detection of RSNOs. We report here results from a detailed mechanistic and kinetic study for trapping RSNOs by three distinct phosphine probes, including structural identification of novel intermediates and stability studies under physiological conditions. We further show that a triarylphosphine-thiophenyl ester can be used in the absolute quantification of endogenous GSNO in several cancer cell lines, while retaining the elements of the SNO functional group, using an LC–MS-based assay. Finally, we demonstrate that a common product ion (m/z = 309.0), derived from phosphine–RSNO adducts, can be used for the detection of other low-molecular weight nitrosothiols (LMW-RSNOs) in biological samples. Collectively, these findings establish a platform for the phosphine ligation-based, specific and direct detection of RSNOs in biological samples, a powerful tool for expanding the knowledge of the biology and chemistry of NO(•)-mediated phenomena. American Chemical Society 2013-04-24 2013-05-22 /pmc/articles/PMC3663071/ /pubmed/23614769 http://dx.doi.org/10.1021/ja401565w Text en Copyright © 2013 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) |
spellingShingle | Seneviratne, Uthpala Godoy, Luiz C. Wishnok, John S. Wogan, Gerald N. Tannenbaum, Steven R. Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs |
title | Mechanism-Based Triarylphosphine-Ester Probes for
Capture of Endogenous RSNOs |
title_full | Mechanism-Based Triarylphosphine-Ester Probes for
Capture of Endogenous RSNOs |
title_fullStr | Mechanism-Based Triarylphosphine-Ester Probes for
Capture of Endogenous RSNOs |
title_full_unstemmed | Mechanism-Based Triarylphosphine-Ester Probes for
Capture of Endogenous RSNOs |
title_short | Mechanism-Based Triarylphosphine-Ester Probes for
Capture of Endogenous RSNOs |
title_sort | mechanism-based triarylphosphine-ester probes for
capture of endogenous rsnos |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663071/ https://www.ncbi.nlm.nih.gov/pubmed/23614769 http://dx.doi.org/10.1021/ja401565w |
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