A modular DNA scaffold to study protein-protein interactions at single-molecule resolution

  • Nat Nanotechnol. 2019 Oct;14(10):988-993. doi: 10.1038/s41565-019-0542-7.
Dorota Kostrz  1  2 ,  Hannah K Wayment-Steele  3 ,  Jing L Wang  4 ,  Maryne Follenfant  1 ,  Vijay S Pande  5 ,  Terence R Strick  6  7  8 ,  Charlie Gosse  9  10
Affiliations
  • 1. Ecole Normale Supérieure, Institut de Biologie de l'Ecole Normale Supérieure (IBENS) CNRS, INSERM, PSL Research University, Paris, France.
  • 2. Laboratoire de Photonique et de Nanostructures, LPN-CNRS, Marcoussis, France.
  • 3. Department of Chemistry, Stanford University, Stanford, USA.
  • 4. Institut Jacques Monod, CNRS, Université Paris Diderot, Université de Paris, Paris, France.
  • 5. Department of Bioengineering, Stanford University, Stanford, USA.
  • 6. Ecole Normale Supérieure, Institut de Biologie de l'Ecole Normale Supérieure (IBENS) CNRS, INSERM, PSL Research University, Paris, France. [email protected].
  • 7. Institut Jacques Monod, CNRS, Université Paris Diderot, Université de Paris, Paris, France. [email protected].
  • 8. Programme Equipe Labellisée, Ligue Nationale Contre le Cancer, Paris, France. [email protected].
  • 9. Ecole Normale Supérieure, Institut de Biologie de l'Ecole Normale Supérieure (IBENS) CNRS, INSERM, PSL Research University, Paris, France. [email protected].
  • 10. Laboratoire de Photonique et de Nanostructures, LPN-CNRS, Marcoussis, France. [email protected].
PMID: 31548690 DOI: 10.1038/s41565-019-0542-7
Abstract

The residence time of a drug on its target has been suggested as a more pertinent metric of therapeutic efficacy than the traditionally used affinity constant. Here, we introduce junctured-DNA tweezers as a generic platform that enables real-time observation, at the single-molecule level, of biomolecular interactions. This tool corresponds to a double-strand DNA scaffold that can be nanomanipulated and on which proteins of interest can be engrafted thanks to widely used genetic tagging strategies. Thus, junctured-DNA tweezers allow a straightforward and robust access to single-molecule force spectroscopy in drug discovery, and more generally in biophysics. Proof-of-principle experiments are provided for the rapamycin-mediated association between FKBP12 and FRB, a system relevant in both medicine and chemical biology. Individual interactions were monitored under a range of applied forces and temperatures, yielding after analysis the characteristic features of the energy profile along the dissociation landscape.

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