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  • 标题:Fabrication of molecular tension probes
  • 本地全文:下载
  • 作者:Sung Bae Kim ; Rika Fujii
  • 期刊名称:MethodsX
  • 印刷版ISSN:2215-0161
  • 电子版ISSN:2215-0161
  • 出版年度:2016
  • 卷号:3
  • 页码:261-267
  • DOI:10.1016/j.mex.2016.03.008
  • 语种:English
  • 出版社:Elsevier
  • 摘要:Abstract 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 determining molecular tension appended by ligand-activated PPIs. The summary of the method is: • Molecular tension appended by protein–protein interactions (PPI) is visualized with a luciferase. • Estrogen activities are quantitatively illuminated with the molecular tension probes. • Full-length Renilla luciferase enhances the optical intensities after bending by PPI.
  • 关键词:Luciferase;Bioluminescence;Molecular tension;Protein–protein interactions (PPI);Renilla luciferase, Bioluminescence imaging (BLI);Estrogen receptor
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