Unraveling the interaction mechanism between the genetic disease drug Ataluren and eukaryotic codon recognition factor 1/ human serum albumin through spectroscopic and nuclear magnetic resonance approaches.

PTC124 (Ataluren) is a small molecule drug for the treatment of genetic diseases that has been shown to promote early termination codon read-through. In this study, the binding model of PTC124 to human serum albumin and eukaryotic stop codon recognition factor (eRF1) was established in vitro. In the cell model, the read-through effect of PTC124 on the stop codon was confirmed. Fluorescence analysis showed that PTC124 could spontaneously bind eRF1 and HSA, the binding constants were 3.132 ± 0.643 × 104 and 2.963 ± 0.218 × 105 L/mol (298 K), respectively. Thermodynamic analysis indicated that the binding to eRF1 was driven mainly by hydrophobic interactions, hydrogen bonding, and van der Waals forces, whereas the binding to HSA was driven primarily by hydrogen bonding and van der Waals forces. Structural analysis revealed perturbations in the secondary structure of both HSA and eRF1 induced by PTC124, while the overall native folds of the proteins were maintained. In addition, a binding model of the interaction between PTC124 and eRF1/HSA was established by molecular simulation and STD-NMR. The binding model described in this paper explains the interaction mechanism between PTC124 and its utility and transport targets at the molecular level, which is helpful for the design and synthesis of new molecules related to PTC124 structure and based on its mechanism of action.