Critical assessment of selenourea as an efficient small molecule fluorescence quenching probe to monitor protein dynamics

Organoselenium compounds have recently been the experimentalists' delight due to their broad applications in organic synthesis, medicinal chemistry, and materials science. Selenium atom replacement of the carbonyl oxygen of the urea moiety dramatically reduces the HOMO-LUMO gap and oxidation potential, which completely changes the physicochemical properties of selenocarbonyl compounds. To our surprise, the photophysics and utility of a simple molecule such as selenourea (SeU) have not been explored in detail, which persuaded us to investigate its role in excited state processes. The steady-state emission, temperature-dependent time-correlated single photon counting, and femtosecond fluorescence upconversion experimental results confirmed that SeU significantly enhances the fluorescence quenching through a photoinduced electron transfer (PET) mechanism with an similar to 10 ps ultrafast intrinsic PET lifetime component which is mostly absent in thiourea (TU). A wide range of fluorophores, based on their different redox abilities and fluorescence lifetimes covering a broad spectral window (lambda ex: 390-590 nm and lambda em: 490-690 nm), were chosen to validate the proof of the concept. It was extended to tetramethylrhodamine (TMR)-5-maleimide labeled lysozyme protein, where we observed significant fluorescence quenching in the presence of SeU. The present work emphasizes that the high quenching efficiency with an ultrafast PET process, reduced orbital energy gap, and higher negative free energy change of the electron transfer reaction are the representative characteristics of selenourea or selenoamides to enable them as potential surrogates of thioamides or oxoamides quenching probes to monitor protein conformational changes and dynamics. This work unravels that reduced Delta E and higher -ve Delta G of the electron transfer reaction are crucial for an ultrafast intrinsic photoinduced electron transfer (PET) process that enables selenourea (SeU) as an efficient fluorescence quenching probe.

Automated summary

Organoselenium compounds have broad applications in organic synthesis, medicinal chemistry, and materials science. A study discovered that selenourea (SeU) significantly enhances fluorescence quenching through a photoinduced electron transfer mechanism and has a fast intrinsic PET lifetime. This research reveals the potential applications of selenourea.