IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v11y2020i1d10.1038_s41467-020-15465-9.html
   My bibliography  Save this article

A HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues

Author

Listed:
  • Jaime Andrés Rivas-Pardo

    (Columbia University
    Universidad Mayor)

  • Yong Li

    (University of Muenster)

  • Zsolt Mártonfalvi

    (Semmelweis University)

  • Rafael Tapia-Rojo

    (Columbia University)

  • Andreas Unger

    (University of Muenster)

  • Ángel Fernández-Trasancos

    (Centro Nacional de Investigaciones Cardiovasculares (CNIC))

  • Elías Herrero-Galán

    (Centro Nacional de Investigaciones Cardiovasculares (CNIC))

  • Diana Velázquez-Carreras

    (Centro Nacional de Investigaciones Cardiovasculares (CNIC))

  • Julio M. Fernández

    (Columbia University)

  • Wolfgang A. Linke

    (University of Muenster)

  • Jorge Alegre-Cebollada

    (Centro Nacional de Investigaciones Cardiovasculares (CNIC))

Abstract

Single-molecule methods using recombinant proteins have generated transformative hypotheses on how mechanical forces are generated and sensed in biological tissues. However, testing these mechanical hypotheses on proteins in their natural environment remains inaccesible to conventional tools. To address this limitation, here we demonstrate a mouse model carrying a HaloTag-TEV insertion in the protein titin, the main determinant of myocyte stiffness. Using our system, we specifically sever titin by digestion with TEV protease, and find that the response of muscle fibers to length changes requires mechanical transduction through titin’s intact polypeptide chain. In addition, HaloTag-based covalent tethering enables examination of titin dynamics under force using magnetic tweezers. At pulling forces

Suggested Citation

  • Jaime Andrés Rivas-Pardo & Yong Li & Zsolt Mártonfalvi & Rafael Tapia-Rojo & Andreas Unger & Ángel Fernández-Trasancos & Elías Herrero-Galán & Diana Velázquez-Carreras & Julio M. Fernández & Wolfgang , 2020. "A HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15465-9
    DOI: 10.1038/s41467-020-15465-9
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-020-15465-9
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-020-15465-9?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
    ---><---

    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:11:y:2020:i:1:d:10.1038_s41467-020-15465-9. 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.