A thiol probe for measuring unfolded protein load and proteostasis in cells.
A thiol probe for measuring unfolded protein load and proteostasis in cells.
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DOI：
10.1038/s41467-017-00203-5
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发表时间：
2017-09-07
影响因子：
16.6
通讯作者：
Hatters DM
中科院分区：
文献类型：
--
作者：
Chen MZ;Moily NS;Bridgford JL;Wood RJ;Radwan M;Smith TA;Song Z;Tang BZ;Tilley L;Xu X;Reid GE;Pouladi MA;Hong Y;Hatters DM
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. Proteostasis is maintained through a number of molecular mechanisms, some of which function to protect the folded state of proteins. Here the authors demonstrate the use of TPE-MI in a fluorigenic dye assay for the quantitation of unfolded proteins that can be used to assess proteostasis on a cellular or proteome scale.
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影响因子：
9.9
作者：
通讯作者：
--
DOI：
10.1073/pnas.1600459113
发表时间：
2016-02-23
影响因子：
11.1
作者：
Ismail, Hanafy M.;Barton, Victoria;Ward, Stephen A.
通讯作者：
Ward, Stephen A.
影响因子：
7.2
作者：
Hutt, Darren M.;Balch, William E.
通讯作者：
Balch, William E.
影响因子：
11.4
作者：
Ivanov, Pavel V.;Gehring, Niels H.;Kulozik, Andreas E.
通讯作者：
Kulozik, Andreas E.
影响因子：
10.5
作者：
ABRAVAYA, K;PHILLIPS, B;MORIMOTO, RI
通讯作者：
MORIMOTO, RI