An ALuc-Based Molecular Tension Probe for Sensing Intramolecular Protein–Protein Interactions

  • Sung-Bae KimEmail author
  • Ryo Nishihara
  • Koji Suzuki
Part of the Methods in Molecular Biology book series (MIMB, volume 1461)


Optical imaging of protein–protein interactions (PPIs) facilitates comprehensive elucidation of intracellular molecular events. The present protocol demonstrates an optical measure for visualizing molecular tension triggered by any PPI in mammalian cells. A unique design of single-chain probes was fabricated, in which a full-length artificial luciferase (ALuc®) was sandwiched between two model proteins of interest, e.g., FKBP and FRB. A molecular tension probe comprising ALuc23 greatly enhances the bioluminescence in response to varying concentrations of rapamycin, and named “tension probe (TP).” The basic probe design can be further modified towards eliminating the C-terminal end of ALuc and was found to improve signal-to-background ratios, named “combinational probe.” TPs may become an important addition to the tool box of bioassays in the determination of protein dynamics of interest in mammalian cells.

Key words

Bioluminescence Protein–protein interactions Tension probe Bioluminescent imaging Combinational probe 



This work was partly supported by JSPS KAKENHI Grants Number 26288088, 16K14051, and 15KK0029.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST)TsukubaJapan
  2. 2.Department of Applied Chemistry, Faculty of Science and TechnologyKeio UniversityYokohamaJapan

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