Methods : [time-resolved] room temperature crystallography

Room temperature crystallography gives better insights on physiological protein dynamics than cryocrystallography

Contact : Sylvain Engilberge & Antoine Royant

Time-resolved macromolecular crystallography (TR-MX) at synchrotrons has long been limited to the use of the Laue diffraction technique and was only successful for a handful of biological systems. The recent advent of X-ray free-electron lasers has rejuvenated the field by fostering the development of serial crystallography, which consists in composing a complete data set from single diffraction images obtained from tens of thousands of microcrystals passing through a pulsed X-ray beam. The associated development of crystal injection techniques, coupled to the advent of faster, noiseless X-ray detectors, has paved the way for the revival of TR-MX at synchrotrons, starting with the generalisation of room temperature MX. We use and develop methods in TR-MX at synchrotrons and XFELs, including the development of time-resolved spectroscopy at the icOS Lab.

Selected publications :

1. Aumonier S, Engilberge S, Caramello C, Gotthard G, Leonard GA, Mueller-Dieckmann C, Royant A (2022) Slow protein dynamics probed by time-resolved oscillation crystallography at room temperature. IUCrJ 9, 756-767 (2022)

2. Maestre-Reyna M, Yang CH, Nango E, Huang WC, Putu EPGN, Wu WJ, Wang PH, Franz-Badur S, Saft M, Emmerich HJ, Wu HY, Lee CC, Huang KF, Chang YK, Liao JH, Weng JH, Gad W, Chang CW, Pang A, Sugahara M, Owada S, Hosokawa Y, Joti Y, Yamashita A, Tanaka R, Tanaka T, Fangjia L, Tono K, Hsu KC, Kiontke S, Schapiro I, Spadaccini R, Royant A, Yamamoto J, Iwata S, Essen LO, Bessho Y, Tsai MD. Serial crystallography captures dynamic control of sequential electron and proton transfer events in a flavoenzyme. Nat. Chem. 14, 677-685 (2022)

3. Ultrafast structural changes within a photosynthetic reaction centre. Dods R, Båth P, Morozov D, Gagnér VA, Arnlund D, Luk HL, Kübel J, Maj M, Vallejos A, Wickstrand C, Bosman R, Beyerlein KR, Nelson G, Liang M, Milathianaki D, Robinson J, Harimoorthy R, Berntsen P, Malmerberg E, Johansson L, Andersson R, Carbajo S, Claesson E, Conrad CE, Dahl P, Hammarin G, Hunter MS, Li C, Lisova S, Royant A, Safari C, Sharma A, Williams GJ, Yefanov O, Westenhoff S, Davidsson J, DePonte DP, Boutet S, Barty A, Katona G, Groenhof G, Brändén G, Neutze R (2021) Nature 589, 310-314

4. Millisecond time-resolved serial oscillation crystallography of a blue-light photoreceptor at a synchrotron. Aumonier S, Santoni G, Gotthard G, von Stetten D, Leonard GA, Royant A (2020) IUCrJ 7, 728-736

5. Specific radiation damage is a lesser concern at room temperature. Gotthard G, Aumonier S, De Sanctis D, Leonard G, von Stetten D, Royant A (2019) IUCrJ 6, 665-680