A thiol probe for measuring unfolded protein load and proteostasis in cells

When proteostasis becomes unbalanced, unfolded proteins can accumulate and aggregate. Here we report that the dye, tetraphenylethene maleimide (TPE-MI) can be used to measure cellular unfolded protein load. TPE-MI fluorescence is activated upon labelling free cysteine thiols, normally buried in the...

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Autores principales: Chen, Moore Z., Moily, Nagaraj S., Bridgford, Jessica L., Wood, Rebecca J., Radwan, Mona, Smith, Trevor A., Song, Zhegang, Tang, Ben Zhong, Tilley, Leann, Xu, Xiaohong, Reid, Gavin E., Pouladi, Mahmoud A., Hong, Yuning, Hatters, Danny M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
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Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589734/
https://www.ncbi.nlm.nih.gov/pubmed/28883394
http://dx.doi.org/10.1038/s41467-017-00203-5
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author Chen, Moore Z.
Moily, Nagaraj S.
Bridgford, Jessica L.
Wood, Rebecca J.
Radwan, Mona
Smith, Trevor A.
Song, Zhegang
Tang, Ben Zhong
Tilley, Leann
Xu, Xiaohong
Reid, Gavin E.
Pouladi, Mahmoud A.
Hong, Yuning
Hatters, Danny M.
author_facet Chen, Moore Z.
Moily, Nagaraj S.
Bridgford, Jessica L.
Wood, Rebecca J.
Radwan, Mona
Smith, Trevor A.
Song, Zhegang
Tang, Ben Zhong
Tilley, Leann
Xu, Xiaohong
Reid, Gavin E.
Pouladi, Mahmoud A.
Hong, Yuning
Hatters, Danny M.
author_sort Chen, Moore Z.
collection PubMed
description When proteostasis becomes unbalanced, unfolded proteins can accumulate and aggregate. Here we report that the dye, tetraphenylethene maleimide (TPE-MI) can be used to measure cellular unfolded protein load. TPE-MI fluorescence is activated upon labelling free cysteine thiols, normally buried in the core of globular proteins that are exposed upon unfolding. Crucially TPE-MI does not become fluorescent when conjugated to soluble glutathione. We find that TPE-MI fluorescence is enhanced upon reaction with cellular proteomes under conditions promoting accumulation of unfolded proteins. TPE-MI reactivity can be used to track which proteins expose more cysteine residues under stress through proteomic analysis. We show that TPE-MI can report imbalances in proteostasis in induced pluripotent stem cell models of Huntington disease, as well as cells transfected with mutant Huntington exon 1 before the formation of visible aggregates. TPE-MI also detects protein damage following dihydroartemisinin treatment of the malaria parasites Plasmodium falciparum. TPE-MI therefore holds promise as a tool to probe proteostasis mechanisms in disease.
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spelling pubmed-55897342017-09-11 A thiol probe for measuring unfolded protein load and proteostasis in cells Chen, Moore Z. Moily, Nagaraj S. Bridgford, Jessica L. Wood, Rebecca J. Radwan, Mona Smith, Trevor A. Song, Zhegang Tang, Ben Zhong Tilley, Leann Xu, Xiaohong Reid, Gavin E. Pouladi, Mahmoud A. Hong, Yuning Hatters, Danny M. Nat Commun Article When proteostasis becomes unbalanced, unfolded proteins can accumulate and aggregate. Here we report that the dye, tetraphenylethene maleimide (TPE-MI) can be used to measure cellular unfolded protein load. TPE-MI fluorescence is activated upon labelling free cysteine thiols, normally buried in the core of globular proteins that are exposed upon unfolding. Crucially TPE-MI does not become fluorescent when conjugated to soluble glutathione. We find that TPE-MI fluorescence is enhanced upon reaction with cellular proteomes under conditions promoting accumulation of unfolded proteins. TPE-MI reactivity can be used to track which proteins expose more cysteine residues under stress through proteomic analysis. We show that TPE-MI can report imbalances in proteostasis in induced pluripotent stem cell models of Huntington disease, as well as cells transfected with mutant Huntington exon 1 before the formation of visible aggregates. TPE-MI also detects protein damage following dihydroartemisinin treatment of the malaria parasites Plasmodium falciparum. TPE-MI therefore holds promise as a tool to probe proteostasis mechanisms in disease. Nature Publishing Group UK 2017-09-07 /pmc/articles/PMC5589734/ /pubmed/28883394 http://dx.doi.org/10.1038/s41467-017-00203-5 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Chen, Moore Z.
Moily, Nagaraj S.
Bridgford, Jessica L.
Wood, Rebecca J.
Radwan, Mona
Smith, Trevor A.
Song, Zhegang
Tang, Ben Zhong
Tilley, Leann
Xu, Xiaohong
Reid, Gavin E.
Pouladi, Mahmoud A.
Hong, Yuning
Hatters, Danny M.
A thiol probe for measuring unfolded protein load and proteostasis in cells
title A thiol probe for measuring unfolded protein load and proteostasis in cells
title_full A thiol probe for measuring unfolded protein load and proteostasis in cells
title_fullStr A thiol probe for measuring unfolded protein load and proteostasis in cells
title_full_unstemmed A thiol probe for measuring unfolded protein load and proteostasis in cells
title_short A thiol probe for measuring unfolded protein load and proteostasis in cells
title_sort thiol probe for measuring unfolded protein load and proteostasis in cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589734/
https://www.ncbi.nlm.nih.gov/pubmed/28883394
http://dx.doi.org/10.1038/s41467-017-00203-5
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