An Aluc-based molecular tension probe for sensing intramolecular protein–Protein interactions

Sung Bae Kim, Ryo Nishihara, Koji Suzuki

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages183-193
Number of pages11
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1461
ISSN (Print)1064-3745

Keywords

  • Bioluminescence
  • Bioluminescent imaging
  • Combinational probe
  • Protein–protein interactions
  • Tension probe

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Kim, S. B., Nishihara, R., & Suzuki, K. (2016). An Aluc-based molecular tension probe for sensing intramolecular protein–Protein interactions. In Methods in Molecular Biology (pp. 183-193). (Methods in Molecular Biology; Vol. 1461). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3813-1_15