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Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
The reversible modification of cysteine residues through thioester formation with palmitate (protein S-palmitoylation) is a prevalent chemical modification that regulates the function, localization, and stability of many proteins. Current methods for monitoring the “erasers” of S-palmitoylation, acy...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5848818/ https://www.ncbi.nlm.nih.gov/pubmed/29568422 http://dx.doi.org/10.1039/c7sc02805a |
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author | Beck, Michael W. Kathayat, Rahul S. Cham, Candace M. Chang, Eugene B. Dickinson, Bryan C. |
author_facet | Beck, Michael W. Kathayat, Rahul S. Cham, Candace M. Chang, Eugene B. Dickinson, Bryan C. |
author_sort | Beck, Michael W. |
collection | PubMed |
description | The reversible modification of cysteine residues through thioester formation with palmitate (protein S-palmitoylation) is a prevalent chemical modification that regulates the function, localization, and stability of many proteins. Current methods for monitoring the “erasers” of S-palmitoylation, acyl-protein thioesterases (APTs), rely on destructive proteomic methods or “turn-on” probes, precluding deployment in heterogeneous samples such as primary tissues. To address these challenges, we present the design, synthesis, and biological evaluation of Ratiometric Depalmitoylation Probes (RDPs). RDPs respond to APTs with a robust ratiometric change in fluorescent signal both in vitro and in live cells. Moreover, RDPs can monitor endogenous APT activities in heterogeneous primary human tissues such as colon organoids, presaging the utility of these molecules in uncovering novel roles for APTs in metabolic regulation. |
format | Online Article Text |
id | pubmed-5848818 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-58488182018-03-22 Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues Beck, Michael W. Kathayat, Rahul S. Cham, Candace M. Chang, Eugene B. Dickinson, Bryan C. Chem Sci Chemistry The reversible modification of cysteine residues through thioester formation with palmitate (protein S-palmitoylation) is a prevalent chemical modification that regulates the function, localization, and stability of many proteins. Current methods for monitoring the “erasers” of S-palmitoylation, acyl-protein thioesterases (APTs), rely on destructive proteomic methods or “turn-on” probes, precluding deployment in heterogeneous samples such as primary tissues. To address these challenges, we present the design, synthesis, and biological evaluation of Ratiometric Depalmitoylation Probes (RDPs). RDPs respond to APTs with a robust ratiometric change in fluorescent signal both in vitro and in live cells. Moreover, RDPs can monitor endogenous APT activities in heterogeneous primary human tissues such as colon organoids, presaging the utility of these molecules in uncovering novel roles for APTs in metabolic regulation. Royal Society of Chemistry 2017-11-01 2017-09-11 /pmc/articles/PMC5848818/ /pubmed/29568422 http://dx.doi.org/10.1039/c7sc02805a Text en This journal is © The Royal Society of Chemistry 2017 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0) |
spellingShingle | Chemistry Beck, Michael W. Kathayat, Rahul S. Cham, Candace M. Chang, Eugene B. Dickinson, Bryan C. Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues |
title | Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
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title_full | Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
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title_fullStr | Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
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title_full_unstemmed | Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
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title_short | Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
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title_sort | michael addition-based probes for ratiometric fluorescence imaging of protein s-depalmitoylases in live cells and tissues |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5848818/ https://www.ncbi.nlm.nih.gov/pubmed/29568422 http://dx.doi.org/10.1039/c7sc02805a |
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