Protein Structure Data and Precise Predictions Pave the Way for Molecular Design

X-ray crystallography remains a powerful technique for determining protein three-dimensional structures with high spatial resolution and high throughput, even though cryo-electron microscopy has become widely used. In conventional X-ray crystallography, most obtained structures were static. However, serial femtosecond crystallography enables the visualization of dynamic structures with high temporal and spatial resolution by combining reaction triggers such as light excitation and substrate mixing. In addition, we have recently established and begun operating an X-ray crystallography experimental station at the fourth-generation synchrotron radiation facility NanoTerasu. In this article, we introduce recent topics in X-ray protein crystallography, including not only experimental advances but also widely used protein structure prediction and molecular design tools.