Kim Sung-Bae, Fujii Rika
Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki, 305-8569, Japan.
Methods Mol Biol. 2016;1461:175-82. 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.
一种独特的生物发光成像探针被引入，用于照亮由感兴趣的蛋白质-蛋白质相互作用（PPI）附加的分子张力。一个全长荧光素酶通过最小的柔性接头夹在两个感兴趣的蛋白质之间。配体激活的PPI会给夹在中间的荧光素酶附加分子内张力，以定量方式提高或降低酶活性。该方法指导构建用于配体激活的PPI的新系列生物测定法。
