IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v8y2017i1d10.1038_s41467-017-00203-5.html
   My bibliography  Save this article

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
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-017-00203-5
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-017-00203-5?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    Citations

    Citations are extracted by the CitEc Project, subscribe to its RSS feed for this item.
    as


    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 & Eren 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.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-00203-5. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.