Research
Studying molecular mechanisms behind the protein function is a challenging task due to the very high spatiotemporal resolution needed for a detailed understanding of the 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.
Our group builds particularly on the recent experiments that focused on 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 the serial crystallography experiments at the synchrotron, which in combination with a pump laser provides an excellent tool to probe slower protein dynamics at more widely available X-ray sources4,5.
The exciting insights into the bacteriorhodopsin and other model proteins inspire us to apply a similar methodology to relatively poorly understood enzymes and transport proteins. The structural work is extensively complemented by other biochemical and biophysical methods, in particular, supported by collaborations involving 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).