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https://doi.org/10.1038/s41467-017-00203-5
| Title: | A thiol probe for measuring unfolded protein load and proteostasis in cells | Authors: | Chen, M.Z Moily, N.S Bridgford, J.L Wood, R.J Radwan, M Smith, T.A Song, Z Tang, B.Z Tilley, L Xu, X Reid, G.E Pouladi, M.A Hong, Y Hatters, D.M |
Keywords: | beta lactoglobulin cysteine dihydroartemisinin dye enolase globular protein glutathione huntingtin peroxiredoxin 3 protein proteome tetraphenylethene maleimide thiol ubiquitin unclassified drug artemisinin derivative epoxomicin fluorescent dye maleimide maleimide derivative mutant protein oligopeptide thiol derivative tunicamycin cells and cell components dye nervous system disorder parasite protein proteomics reaction kinetics thiol Article exon human Huntington chorea induced pluripotent stem cell nonhuman Plasmodium falciparum protein folding protein homeostasis protein unfolding animal cells chemistry drug effects HEK293 cell line HeLa cell line malaria metabolism molecular probe mouse parasite parasitology protein folding solubility Plasmodium falciparum Animals Artemisinins Cells Cysteine Fluorescent Dyes HEK293 Cells HeLa Cells Humans Huntingtin Protein Malaria Maleimides Mice Molecular Probes Mutant Proteins Oligopeptides Parasites Protein Folding Proteome Proteostasis Solubility Sulfhydryl Compounds Tunicamycin |
Issue Date: | 2017 | Publisher: | Nature Publishing Group | Citation: | Chen, M.Z, Moily, N.S, Bridgford, J.L, Wood, R.J, Radwan, M, Smith, T.A, Song, Z, Tang, B.Z, Tilley, L, Xu, X, Reid, G.E, Pouladi, M.A, Hong, Y, Hatters, D.M (2017). A thiol probe for measuring unfolded protein load and proteostasis in cells. Nature Communications 8 (1) : 474. ScholarBank@NUS Repository. https://doi.org/10.1038/s41467-017-00203-5 | Rights: | Attribution 4.0 International | Abstract: | 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. © 2017 The Author(s). | Source Title: | Nature Communications | URI: | https://scholarbank.nus.edu.sg/handle/10635/178580 | ISSN: | 2041-1723 | DOI: | 10.1038/s41467-017-00203-5 | Rights: | Attribution 4.0 International |
| Appears in Collections: | Staff Publications Elements |
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