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

Sung Bae Kim, Ryo Nishihara, Koji Suzuki

研究成果: Chapter

抄録

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.

本文言語English
ホスト出版物のタイトルMethods in Molecular Biology
出版社Humana Press Inc.
ページ183-193
ページ数11
DOI
出版ステータスPublished - 2016

出版物シリーズ

名前Methods in Molecular Biology
1461
ISSN(印刷版)1064-3745

ASJC Scopus subject areas

  • 分子生物学
  • 遺伝学

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