A FlAsH-Based Cross-Linker to Study Protein Interactions in Living Cells

Authors

  • Dr. Anna Rutkowska,

    1. Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Dr. Christian H. Haering,

    Corresponding author
    1. Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
    • Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • Priv.-Doz. Dr. Carsten Schultz

    Corresponding author
    1. Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
    • Cell Biology & Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg (Germany)
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  • We thank the Advanced Light Microscopy Facility at EMBL, Heike Stichnoth for providing cells, Andrea Erlbruch and Alen Piljic for providing initial PKA, DAPK1, and calmodulin constructs, Birgit Koch for providing the anti-GFP antibody, Sarah Sternberger for help during experiments with purified protein, and Stephen R. Adams (UCSD) for discussions. A.R. is a fellow of the Peter und Traudl Engelhorn-Stiftung and the EMBL Interdisciplinary Postdoc Programme (EIPOD). FlAsH=fluorescein-based arsenical hairpin binder.

Abstract

original image

As you like it: xCrAsH, a dimeric derivative of the arsenical compound FlAsH, enables the highly specific, covalent cross-linking of two proteins containing a 12 amino acid peptide tag. This inducible and (by addition of dithiols) reversible system can be used to detect and manipulate protein–protein interactions both in vitro and in living cells (see picture).

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