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A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors
Phosphodiesterase type 5 (PDE5), a cyclic nucleotide phosphodiesterase, controls the duration of the cyclic guanosine monophosphate (cGMP) signal by hydrolyzing cGMP to GMP. Inhibiting the activity of PDE5A has proven to be an effective strategy for treating pulmonary arterial hypertension and erect...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9968969/ https://www.ncbi.nlm.nih.gov/pubmed/36860644 http://dx.doi.org/10.3389/fchem.2023.1097027 |
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author | Ma, Yufeng Zhang, Fengsen Zhong, Yijing Huang, Yongchun Yixizhuoma, Jia, Qiangqiang Zhang, Shoude |
author_facet | Ma, Yufeng Zhang, Fengsen Zhong, Yijing Huang, Yongchun Yixizhuoma, Jia, Qiangqiang Zhang, Shoude |
author_sort | Ma, Yufeng |
collection | PubMed |
description | Phosphodiesterase type 5 (PDE5), a cyclic nucleotide phosphodiesterase, controls the duration of the cyclic guanosine monophosphate (cGMP) signal by hydrolyzing cGMP to GMP. Inhibiting the activity of PDE5A has proven to be an effective strategy for treating pulmonary arterial hypertension and erectile dysfunction. Current enzymatic activity assay methods for PDE5A mainly use fluorescent or isotope-labeled substrates, which are expensive and inconvenient. Here, we developed an LC/MS-based enzymatic activity assay for PDE5A without labeling, which detects the enzymatic activity of PDE5A by quantifying the substrate cGMP and product GMP at a concentration of 100 nM. The accuracy of this method was verified by a fluorescently labeled substrate. Moreover, a new inhibitor of PDE5A was identified by this method and virtual screening. It inhibited PDE5A with an IC(50) value of 870 nM. Overall, the proposed strategy provides a new method for screening PDE5A inhibitors. |
format | Online Article Text |
id | pubmed-9968969 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-99689692023-02-28 A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors Ma, Yufeng Zhang, Fengsen Zhong, Yijing Huang, Yongchun Yixizhuoma, Jia, Qiangqiang Zhang, Shoude Front Chem Chemistry Phosphodiesterase type 5 (PDE5), a cyclic nucleotide phosphodiesterase, controls the duration of the cyclic guanosine monophosphate (cGMP) signal by hydrolyzing cGMP to GMP. Inhibiting the activity of PDE5A has proven to be an effective strategy for treating pulmonary arterial hypertension and erectile dysfunction. Current enzymatic activity assay methods for PDE5A mainly use fluorescent or isotope-labeled substrates, which are expensive and inconvenient. Here, we developed an LC/MS-based enzymatic activity assay for PDE5A without labeling, which detects the enzymatic activity of PDE5A by quantifying the substrate cGMP and product GMP at a concentration of 100 nM. The accuracy of this method was verified by a fluorescently labeled substrate. Moreover, a new inhibitor of PDE5A was identified by this method and virtual screening. It inhibited PDE5A with an IC(50) value of 870 nM. Overall, the proposed strategy provides a new method for screening PDE5A inhibitors. Frontiers Media S.A. 2023-02-13 /pmc/articles/PMC9968969/ /pubmed/36860644 http://dx.doi.org/10.3389/fchem.2023.1097027 Text en Copyright © 2023 Ma, Zhang, Zhong, Huang, Yixizhuoma, Jia and Zhang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Chemistry Ma, Yufeng Zhang, Fengsen Zhong, Yijing Huang, Yongchun Yixizhuoma, Jia, Qiangqiang Zhang, Shoude A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title | A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title_full | A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title_fullStr | A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title_full_unstemmed | A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title_short | A label-free LC/MS-based enzymatic activity assay for the detection of PDE5A inhibitors |
title_sort | label-free lc/ms-based enzymatic activity assay for the detection of pde5a inhibitors |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9968969/ https://www.ncbi.nlm.nih.gov/pubmed/36860644 http://dx.doi.org/10.3389/fchem.2023.1097027 |
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