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Review
. 2014 Jul 17;369(1647):20130318.
doi: 10.1098/rstb.2013.0318.

Opportunities and challenges for time-resolved studies of protein structural dynamics at X-ray free-electron lasers

Richard Neutze  1
Affiliations
Review

Opportunities and challenges for time-resolved studies of protein structural dynamics at X-ray free-electron lasers

Richard Neutze. Philos Trans R Soc Lond B Biol Sci. .

Abstract

X-ray free-electron lasers (XFELs) are revolutionary X-ray sources. Their time structure, providing X-ray pulses of a few tens of femtoseconds in duration; and their extreme peak brilliance, delivering approximately 10(12) X-ray photons per pulse and facilitating sub-micrometre focusing, distinguish XFEL sources from synchrotron radiation. In this opinion piece, I argue that these properties of XFEL radiation will facilitate new discoveries in life science. I reason that time-resolved serial femtosecond crystallography and time-resolved wide angle X-ray scattering are promising areas of scientific investigation that will be advanced by XFEL capabilities, allowing new scientific questions to be addressed that are not accessible using established methods at storage ring facilities. These questions include visualizing ultrafast protein structural dynamics on the femtosecond to picosecond time-scale, as well as time-resolved diffraction studies of non-cyclic reactions. I argue that these emerging opportunities will stimulate a renaissance of interest in time-resolved structural biochemistry.

Keywords: X-ray free-electron lasers; serial femtosecond crystallography; structural biology; time-resolved diffraction; time-resolved wide angle X-ray scattering.

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Figures

Figure 1.
Figure 1.
SFX and time-resolved Laue diffraction studies of the photosynthetic reaction centre of Bl. viridis (RCvir). (a) Injection of RCvir microcrystals into an XFEL beam using the Spence microjet. (b) SFX structure of RCvir solved to 3.5 Å resolution. (c) Close-up view of the SFX electron density map near the special pair (P960) and near TyrL162. (d) Similar view as in (c), but of a Laue diffraction electron density map to 2.95 Å resolution. All 2Fobs–Fcalc electron density maps (blue) are contoured at 1.0σ. (e) Difference density (green positive density; red negative density, contoured at 4.0σ) illustrating the structural changes induced by light (the movement of TyrL162 towards the special pair) captured using time-resolved Laue diffraction. These figures are reproduced with permission from [16] (a), [20] (b,c) and [42] (d,e, with modifications).
Figure 2.
Figure 2.
Time-resolved WAXS studies of light-driven structural changes in bacteriorhodopsin. (a) Time-resolved WAXS difference data (ΔS(q,Δt)) as a function of the time-delay (Δt) following photoactivation by a short visible laser pulse. (b) Two difference WAXS basis spectra extracted from spectral decomposition of the data shown in (a) (dots), and the theoretical fits to this data (solid lines), for an intermediate time-scale (black) and slower (red) component of the data. (c) Refined conformational changes in bacteriorhodopsin recovered by a best-fit analysis to the experimental difference WAXS basis spectra shown in (b). These figures are reproduced with permission from [75].
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