Genetically Encoded Molecular Tension Probe for Tracing Protein-Protein Interactions in Mammalian Cells

Sung Bae Kim, Ryo Nishihara, Daniel Citterio, Koji Suzuki

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Optical imaging of protein-protein interactions (PPIs) facilitates comprehensive elucidation of intracellular molecular events. We demonstrate an optical measure for visualizing molecular tension triggered by any PPI in mammalian cells. Twenty-three kinds of candidate designs were fabricated, in which a full-length artificial luciferase (ALuc) was sandwiched between two model proteins of interest, e.g., FKBP and FRB. One of the designs greatly enhanced the bioluminescence in response to varying concentrations of rapamycin. It is confirmed with negative controls that the elevated bioluminescence is solely motivated from the molecular tension. The probe design was further modified toward eliminating the C-terminal end of ALuc and was found to improve signal-to-background ratios, named "a combinational probe". The utilities were elucidated with detailed substrate selectivity, bioluminescence imaging of live cells, and different PPI models. This study expands capabilities of luciferases as a tool for analyses of molecular dynamics and cell signaling in living subjects.

Original languageEnglish
Pages (from-to)354-362
Number of pages9
JournalBioconjugate Chemistry
Volume27
Issue number2
DOIs
Publication statusPublished - 2016 Feb 17

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Molecular Probes
Cells
Proteins
Bioluminescence
Luciferases
Tacrolimus Binding Proteins
Cell signaling
Imaging techniques
Optical Imaging
Sirolimus
Molecular Dynamics Simulation
Molecular dynamics
Substrates

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Genetically Encoded Molecular Tension Probe for Tracing Protein-Protein Interactions in Mammalian Cells. / Kim, Sung Bae; Nishihara, Ryo; Citterio, Daniel; Suzuki, Koji.

In: Bioconjugate Chemistry, Vol. 27, No. 2, 17.02.2016, p. 354-362.

Research output: Contribution to journalArticle

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