Genetically Encoded Molecular Tension Probe for Tracing Protein-Protein Interactions in Mammalian Cells

Bioconjugate Chemistry
Sung Bae KimKoji Suzuki

Abstract

Optical imaging of protein-protein interactions (PPIs) facilitates comprehensive elucidation of intracellular molecular events. We demonstrate an optical measure for visualizing molecular tension triggered by any PPI in mammalian cells. Twenty-three kinds of candidate designs were fabricated, in which a full-length artificial luciferase (ALuc) was sandwiched between two model proteins of interest, e.g., FKBP and FRB. One of the designs greatly enhanced the bioluminescence in response to varying concentrations of rapamycin. It is confirmed with negative controls that the elevated bioluminescence is solely motivated from the molecular tension. The probe design was further modified toward eliminating the C-terminal end of ALuc and was found to improve signal-to-background ratios, named "a combinational probe". The utilities were elucidated with detailed substrate selectivity, bioluminescence imaging of live cells, and different PPI models. This study expands capabilities of luciferases as a tool for analyses of molecular dynamics and cell signaling in living subjects.

References

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Aug 16, 2011·Bioconjugate Chemistry·Sung Bae KimMasaki Torimura
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Nov 9, 2012·Analytical Chemistry·Takeaki OzawaSung Bae Kim
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Nov 20, 2014·Chemical Communications : Chem Comm·Ryo NishiharaKoji Suzuki

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Citations

May 26, 2016·MethodsX·Sung Bae Kim, Rika Fujii
Dec 7, 2018·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Sung Bae KimKoji Suzuki
Dec 7, 2019·Chemical Communications : Chem Comm·Sung Bae KimRamasamy Paulmurugan
Apr 21, 2017·Scientific Reports·Ryo NishiharaSung Bae Kim
Sep 24, 2020·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Sung-Bae Kim, Ramasamy Paulmurugan
Jul 10, 2021·Angewandte Chemie·Yuxin DuanKhalid Salaita

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