Label-free Single-Molecule Quantification of Rapamycin-induced FKBP-FRB Dimerization for Direct Control of Cellular Mechanotransduction

Nano Lett. 2019 Oct 9;19(10):7514-7525. doi: 10.1021/acs.nanolett.9b03364. Epub 2019 Sep 6.

Abstract

Chemically induced dimerization (CID) has been applied to study numerous biological processes and has important pharmacological applications. However, the complex multistep interactions under various physical constraints involved in CID impose a great challenge for the quantification of the interactions. Furthermore, the mechanical stability of the ternary complexes has not been characterized; hence, their potential application in mechanotransduction studies remains unclear. Here, we report a single-molecule detector that can accurately quantify almost all key interactions involved in CID and the mechanical stability of the ternary complex, in a label-free manner. Its application is demonstrated using rapamycin-induced heterodimerization of FRB and FKBP as an example. We revealed the sufficient mechanical stability of the FKBP/rapamycin/FRB ternary complex and demonstrated its utility in the precise switching of talin-mediated force transmission in integrin-based cell adhesions.

Keywords: CID; magnetic tweezers; mechanotransduction; rapamycin; talin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Humans
  • Mechanotransduction, Cellular / drug effects
  • Mice
  • Protein Multimerization / drug effects
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sirolimus / pharmacology*
  • Tacrolimus Binding Protein 1A / chemistry
  • Tacrolimus Binding Protein 1A / metabolism*

Substances

  • Recombinant Proteins
  • Tacrolimus Binding Protein 1A
  • Sirolimus