RESUMEN
One of the most crucial enzymatic cofactors in the human body is tetrahydrobiopterin, which is acquired through biological synthesis and self-regeneration. During this regenerative process, it undergoes oxidation, deprotonation, further oxidation, and subsequent deprotonation, resulting in the formation of quinonoid-dihydrobiopterin, which then undergoes tautomerization to yield dihydrobiopterin. This study presents the thermodynamic and kinetic properties associated with each stage of the regeneration process using theoretical calculations. The redox potentials for oxidation steps and the pKa values for deprotonation steps are determined employing the Born-Haber cycle and the direct change of free energy in implicit solvent models. The redox metabolites are characterized and confirmed from their calculated absorption spectra using the time-dependent density functional theory method. For the tautomerization steps, an IRC calculation is executed, and rate constants are computed using Eyring's Transition State Theory (TST). The tunnelling probability of the H atom during the tautomerization process is incorporated using Wigner's tunnelling correction in the calculation of the rate constant. Notably, we identify the N3 atom as the most probable deprotonation site for H3B+ and predict its geometry based on our calculations. Furthermore, we elucidate the spectral properties of intermediates involved in the regeneration process, highlighting key electronic transitions responsible for their excitations. Our results indicate that each step of tautomerization occurs along vibrational bending modes. We have observed that these tautomerization processes have high activation energies by optimising transition states. Additionally, considering tunnelling correction can significantly affect the reaction rates associated with these processes. These results provide a comprehensive understanding of the thermodynamics and kinetics of the regeneration process of tetrahydrobiopterin, which will help in the modulation of its biological activity.
RESUMEN
Fluorescein, one of the brightest fluorescent dye molecules, is a widely used fluorophore for various applications from biomedicine to industry. The dianionic form of fluorescein is responsible for its high fluorescence quantum yield. Interestingly, the molecule was found to be nonfluorescent in the gas phase. This characteristic is attributed to the photodetachment process, which out-competes the fluorescence emission in the gas phase. In this work, we show that the calculated vertical and adiabatic detachment energies of fluorescein dianion in the gas and solvent phases account for the drastic differences observed in their fluorescence characteristics. The functional dependence of these detachment energies on the dianion's microsolvation was systematically investigated. The performance of different solvent models was also assessed. The higher thermodynamic stability of fluorescein dianion over the monoanion doublet in the solvent phase plays a crucial role in quenching photodetachment and activating the radiative channel with a high fluorescence quantum yield.