Studying small and fast

Studying molecular mechanisms behind the protein function is challenging due to the high spatiotemporal resolution needed for a detailed understanding of complex protein dynamics. The recent development of X-ray Free Electron Laser (XFEL) facilities opens new opportunities for dynamic structural biology. The intense and short 10-​70 fs X-ray pulses enable pump-probe experiments with unprecedented temporal resolution. Time-resolved serial femtosecond crystallography (TR-​SFX) can provide atomic-level details of proteins in action while probing events at femtosecond to millisecond timescales after activation.  

Capturing protein in motion

Our group builds particularly on the recent experiments that focused on the dynamics of bacteriorhodopsin. This work demonstrated the sample-efficient TR-​SFX at XFELs with high-viscosity injectors while providing unique insights into the activation and photocycle dynamics1-3. Bacteriorhodopsin was also used to perform serial crystallography experiments at the synchrotron. Combined with a pump laser, provides an excellent tool to probe slower protein dynamics at more widely available X-ray sources4-5.

Overview of major events during chloride transport by NmHR captured by time-resolved serial X-ray crystallography. For details, check our publication Mous et al. Science 2022.


The exciting insights into bacteriorhodopsin and other model proteins inspire us to apply a similar methodology to poorly understood enzymes and transport proteins. Our structural work is complemented by other biochemical and biophysical methods, supported by collaborations with time-resolved spectroscopy and QM/MM simulations.


1. Nogly, P. et al. Lipidic cubic phase injector is a viable crystal delivery system for time-​resolved serial crystallography. Nat. Commun. 7, 12314 (2016).

2. Nango, E. et al. A three-​dimensional movie of structural changes in bacteriorhodopsin. Science 354, 1552–1557 (2016).

3. Nogly, P. et al. Retinal isomerization in bacteriorhodopsin captured by a femtosecond x-​ray laser. Science 361, (2018).

4. Nogly, P. et al. Lipidic cubic phase serial millisecond crystallography using synchrotron radiation. IUCrJ 2, 168–176 (2015).

5. Weinert, T. et al. Proton uptake mechanism in bacteriorhodopsin captured by serial synchrotron crystallography. Science 365, 61–65 (2019).