Development of time-resolved protein crystallography methods in the microsecond to millisecond time domain

Abstract: Over the last decade, the development of time-resolved serial crystallography (TR-SX) at XFELs and synchrotrons has allowed researchers to study phenomena occurring in proteins on the femtosecond to minute timescale, taking advantage of many technical and methodological breakthroughs. In such an experiment, protein crystals of various sizes are continuously presented to the X-ray beam. Photoactive proteins have naturally been the initial systems to be studied in TR-SX experiments using pump-probe schemes, where the pump is a pulse of visible light. Other reaction initiations through small molecule diffusion are now gaining momentum. During this PhD, a series of biological questions linking protein structures to their functions were tackled, using time-resolved crystallography methods, some of which were developed during this PhD, at synchrotrons, and occasionally XFELs.