Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs
Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO[superscript •]) 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 pathologie...
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| Format: | Article |
| Language: | en_US |
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American Chemical Society (ACS)
2013
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| Online Access: | http://hdl.handle.net/1721.1/82107 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0003-0771-9889 https://orcid.org/0000-0002-2325-552X |
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| author | Wishnok, John S. Wogan, Gerald N. Seneviratne, Uthpala I. Godoy, Luiz Claudio Tannenbaum, Steven Robert |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Wishnok, John S. Wogan, Gerald N. Seneviratne, Uthpala I. Godoy, Luiz Claudio Tannenbaum, Steven Robert |
| author_sort | Wishnok, John S. |
| collection | MIT |
| description | Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO[superscript •]) 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[superscript •]-mediated phenomena. |
| first_indexed | 2024-09-23T16:43:30Z |
| format | Article |
| id | mit-1721.1/82107 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:43:30Z |
| publishDate | 2013 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/821072022-10-03T07:50:06Z Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs Wishnok, John S. Wogan, Gerald N. Seneviratne, Uthpala I. Godoy, Luiz Claudio Tannenbaum, Steven Robert Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemistry Seneviratne, Uthpala I. Godoy, Luiz Claudio Wishnok, John S. Wogan, Gerald N. Tannenbaum, Steven Robert Nitrosothiols (RSNOs) have been proposed as important intermediates in nitric oxide (NO[superscript •]) 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[superscript •]-mediated phenomena. National Institutes of Health (U.S.) (CA26731) Massachusetts Institute of Technology. Center for Environmental Health Sciences (ES002109) Agilent Technologies 2013-11-13T16:59:08Z 2013-11-13T16:59:08Z 2013-04 2013-02 Article http://purl.org/eprint/type/JournalArticle 0002-7863 1520-5126 http://hdl.handle.net/1721.1/82107 Seneviratne, Uthpala, Luiz C. Godoy, John S. Wishnok, Gerald N. Wogan, and Steven R. Tannenbaum. “Mechanism-Based Triarylphosphine-Ester Probes for Capture of Endogenous RSNOs.” Journal of the American Chemical Society 135, no. 20 (May 22, 2013): 7693-7704. © 2013 American Chemical Society https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0003-0771-9889 https://orcid.org/0000-0002-2325-552X en_US http://dx.doi.org/10.1021/ja401565w Journal of the American Chemical Society Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) ACS |
| spellingShingle | Wishnok, John S. Wogan, Gerald N. Seneviratne, Uthpala I. Godoy, Luiz Claudio Tannenbaum, Steven Robert 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 | http://hdl.handle.net/1721.1/82107 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0003-0771-9889 https://orcid.org/0000-0002-2325-552X |
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