Fabrication of Molecular Strain Probes for Illuminating Protein-Protein Interactions

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1461; p. 175
• Main Authors: Kim, Sung-Bae, Fujii, Rika
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Animals; Cercopithecus aethiops; COS Cells; Ligands; Luciferases - chemistry; Luciferases - genetics; Luciferases - metabolism; Luminescent Agents - chemistry; Luminescent Agents - metabolism; Luminescent Measurements - methods; Molecular Imaging - methods; Molecular Probes - genetics; Molecular Probes - metabolism; Protein Conformation; Protein Interaction Mapping - methods; Structure-Activity Relationship; Molecular tension; Bioluminescence; Protein–protein interactions; Luciferase
• DOI: 10.1007/978-1-4939-3813-1_14

A unique bioluminescent imaging probe is introduced for illuminating molecular tension appended by protein-protein interactions (PPIs) of interest. A full-length luciferase is sandwiched between two proteins of interest via minimal flexible linkers. The ligand-activated PPIs append intramolecular tension to the sandwiched luciferase, boosting or dropping the enzymatic activity in a quantitative manner. This method guides construction of a new lineage of bioassays for ligand-activated PPIs.