A multicrystal diffraction data-collection approach for studying structural dynamics with millisecond temporal resolution

作者：Schubert Robin; Kapis Svetlana; Gicquel Yannig; Bourenkov Gleb; Schneider Thomas R; Heymann Michael; Betzel Christian; Perbandt Markus

来源：IUCrJ, 2016, 3(6): 393-401.

DOI：10.1107/S2052252516016304

• time-resolved crystallography
• fixed target
• multicrystal data collection
• room temperature
• synchrotron radiation
• radiation damage
• structure determination
• protein structure
• X-ray crystallography
• structural biology

Many biochemical processes take place on timescales ranging from femto-seconds to seconds. Accordingly, any time-resolved experiment must be matched to the speed of the structural changes of interest. Therefore, the timescale of interest defines the requirements of the X-ray source, instrumentation and data-collection strategy. In this study, a minimalistic approach for in situ crystallization is presented that requires only a few microlitres of sample solution containing a few hundred crystals. It is demonstrated that complete diffraction data sets, merged from multiple crystals, can be recorded within only a few minutes of beamtime and allow high-resolution structural information of high quality to be obtained with a temporal resolution of 40 ms. Global and site-specific radiation damage can be avoided by limiting the maximal dose per crystal to 400 kGy. Moreover, analysis of the data collected at higher doses allows the time-resolved observation of site-specific radiation damage. Therefore, our approach is well suited to observe structural changes and possibly enzymatic reactions in the low-millisecond regime.

• 出版日期2016-11