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. 2020 Jul 31;2(4):e190053. doi: 10.1148/rycan.2020190053

Figure 4:

Multispectral luciferase complementation. A, The C-terminal fragment of click beetle green (CBG-C) was fused to β transducin repeats-containing proteins (βTrCP). The N-terminal fragment of click beetle red (CBR-N) was fused to IκBα. The N-terminal fragment of click beetle green was fused to β-catenin. The spectral emission of the reconstituted click-beetle luciferases maps to the N-terminal portion. Thus, light produced from the β-catenin/βTrCP interaction (green) can be resolved from the IκBa/βTrCP interaction (red) through spectral unmixing. B–D, The simultaneous quantification of the real-time switching of protein-protein interactions with βTrCP can be measured, depending on the exogenous stimulus or small molecule inhibitor. Data in red indicate IκBα/βTrCP interaction, and data in green indicate β-catenin/βTrCP interaction. (Reprinted, with permission, from reference 45.) GSK3β = glycogen synthase kinese 3 beta, SB-216763 = 3-(2,4-Dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione.

Multispectral luciferase complementation. A, The C-terminal fragment of click beetle green (CBG-C) was fused to β transducin repeats-containing proteins (βTrCP). The N-terminal fragment of click beetle red (CBR-N) was fused to IκBα. The N-terminal fragment of click beetle green was fused to β-catenin. The spectral emission of the reconstituted click-beetle luciferases maps to the N-terminal portion. Thus, light produced from the β-catenin/βTrCP interaction (green) can be resolved from the IκBa/βTrCP interaction (red) through spectral unmixing. B–D, The simultaneous quantification of the real-time switching of protein-protein interactions with βTrCP can be measured, depending on the exogenous stimulus or small molecule inhibitor. Data in red indicate IκBα/βTrCP interaction, and data in green indicate β-catenin/βTrCP interaction. (Reprinted, with permission, from reference 45.) GSK3β = glycogen synthase kinese 3 beta, SB-216763 = 3-(2,4-Dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione.