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

Author

Listed:
  • 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
    The University of Melbourne)

  • Mahmoud A. Pouladi

    (Translational Laboratory in Genetic Medicine, Agency for Science, Technology and Research
    National University of Singapore)

  • Yuning Hong

    (The University of Melbourne
    The University of Melbourne
    La Trobe University)

  • Danny M. Hatters

    (The University of Melbourne)

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.

Suggested Citation

  • 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. Poul, 2017. "A thiol probe for measuring unfolded protein load and proteostasis in cells," Nature Communications, Nature, vol. 8(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-00203-5
    DOI: 10.1038/s41467-017-00203-5
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    Cited by:

    1. Narasaiah Kovuru & Makiko Mochizuki-Kashio & Theresa Menna & Greer Jeffrey & Yuning Hong & Young me Yoon & Zhe Zhang & Peter Kurre, 2024. "Deregulated protein homeostasis constrains fetal hematopoietic stem cell pool expansion in Fanconi anemia," Nature Communications, Nature, vol. 15(1), pages 1-15, December.
    2. Dezerae Cox & Ching-Seng Ang & Nadinath B. Nillegoda & Gavin E. Reid & Danny M. Hatters, 2022. "Hidden information on protein function in censuses of proteome foldedness," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    3. Florisela Herrejon Chavez & Hanzhi Luo & Paolo Cifani & Alli Pine & Karen L. Chu & Suhasini Joshi & Ersilia Barin & Alexandra Schurer & Mandy Chan & Kathryn Chang & Grace Y. Q. Han & Aspen J. Pierson , 2023. "RNA binding protein SYNCRIP maintains proteostasis and self-renewal of hematopoietic stem and progenitor cells," Nature Communications, Nature, vol. 14(1), pages 1-19, December.

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