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

Moore Z. Chen, Nagaraj S. Moily, Jessica L. Bridgford, Rebecca J. Wood, Mona Radwan, Trevor A. Smith, Zhegang Song, Ben Zhong Tang, Leann Tilley, Xiaohong Xu, Gavin E. Reid, Mahmoud A. Pouladi, Yuning Hong () and Danny M. Hatters ()
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Moore Z. Chen: The University of Melbourne
Nagaraj S. Moily: The University of Melbourne
Jessica L. Bridgford: The University of Melbourne
Rebecca J. Wood: The University of Melbourne
Mona Radwan: The University of Melbourne
Trevor A. Smith: The University of Melbourne
Zhegang Song: The Hong Kong University of Science & Technology
Ben Zhong Tang: The Hong Kong University of Science & Technology
Leann Tilley: The University of Melbourne
Xiaohong Xu: Translational Laboratory in Genetic Medicine, Agency for Science, Technology and Research
Gavin E. Reid: The University of Melbourne
Mahmoud A. Pouladi: Translational Laboratory in Genetic Medicine, Agency for Science, Technology and Research
Yuning Hong: The University of Melbourne
Danny M. Hatters: The University of Melbourne

Nature Communications, 2017, vol. 8, issue 1, 1-11

Abstract: 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.

Date: 2017
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DOI: 10.1038/s41467-017-00203-5

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