High-Resolution Protein Structure Determination by Serial Femtosecond Crystallography

@article{Boutet2012HighResolutionPS,
  title={High-Resolution Protein Structure Determination by Serial Femtosecond Crystallography},
  author={S. Boutet and L. Lomb and G. Williams and T. Barends and A. Aquila and R. Doak and U. Weierstall and D. DePonte and J. Steinbrener and R. Shoeman and M. Messerschmidt and A. Barty and T. White and S. Kassemeyer and R. Kirian and M. Seibert and P. Montanez and C. Kenney and R. Herbst and P. Hart and J. Pines and G. Haller and S. Gruner and H. Philipp and M. Tate and M. Hromalik and L. Koerner and N. van Bakel and J. Morse and W. Ghonsalves and D. Arnlund and M. Bogan and C. Caleman and R. Fromme and C. Hampton and M. Hunter and L. Johansson and G. Katona and C. Kupitz and M. Liang and A. V. Martin and K. Nass and L. Redecke and F. Stellato and N. T{\^i}mneanu and D. Wang and N. Zatsepin and D. Schafer and J. Defever and R. Neutze and P. Fromme and J. Spence and H. Chapman and I. Schlichting},
  journal={Science},
  year={2012},
  volume={337},
  pages={362 - 364}
}
Size Matters Less X-ray crystallography is a central research tool for uncovering the structures of proteins and other macromolecules. However, its applicability typically requires growth of large crystals, in part because a sufficient number of molecules must be present in the lattice for the sample to withstand x-ray—induced damage. Boutet et al. (p. 362, published online 31 May) now demonstrate that the intense x-ray pulses emitted by a free-electron laser source can yield data in few enough… Expand
Protein crystallography using x-ray free-electron lasers
Just over 100 years ago, Max von Laue reported the first x-ray diffraction patterns from crystals. Soon after, Lawrence Bragg interpreted diffraction spots. Their work provided the foundation forExpand
Serial protein crystallography in an electron microscope
TLDR
This work presents a serial electron diffraction method, where still diffraction patterns from many protein nanocrystals are rapidly recorded and merged, which minimises radiation damage and only requires a slightly modified standard scanning transmission electron microscope. Expand
De novo protein crystal structure determination from X-ray free-electron laser data
TLDR
X-ray FEL data can be used for de novo protein structure determination, that is, without previous knowledge about the structure, and high-quality diffraction intensities are obtained, resulting in an experimental electron density map good enough for automated building of the protein structure. Expand
Watching Proteins Function with Time-resolved X-ray Crystallography.
TLDR
Time-resolved macromolecular crystallography with 100 ps time resolution at synchrotron X-ray sources is in its mature phase today, particularly for studies of reversible, light-initiated reactions. Expand
Serial femtosecond crystallography: A revolution in structural biology.
TLDR
Structural elucidation while avoiding radiation damage, even at room temperature, can now be achieved in the emerging field of serial femtosecond crystallography (SFX), allowing structural biologists access to previously restricted scientific territory. Expand
Expanding the femtosecond crystallography toolkit
  • S. Gruner
  • Chemistry, Medicine
  • Proceedings of the National Academy of Sciences
  • 2014
TLDR
The latest tool in the crystallographic arsenal, the X-ray free electron laser (XFEL) promises to help surmount challenges, provided efficient ways are found to handle crystals in the demanding XFEL environment, and Cohen et al. (3) show that crystal handling technology developed primarily for use at synchrotron storage ringX-ray sources can also be effectively adapted for use in the XFel environment. Expand
X-ray laser diffraction for structure determination of the rhodopsin-arrestin complex
TLDR
The rhodopsin-arrestin crystal structure solved with SFX represents the first near-atomic resolution structure of a GPCR-Arrestin complex, provides structural insights into understanding of arrestin-mediated GPCRs signaling, and demonstrates the great potential of this SFX-XFEL technology for accelerating crystal structure determination of challenging proteins and protein complexes. Expand
Time-resolved serial crystallography captures high-resolution intermediates of photoactive yellow protein
TLDR
This work used microcrystals of photoactive yellow protein as a model system and obtained high-resolution, time-resolved difference electron density maps of excellent quality with strong features, which open the way to the study of reversible and nonreversible biological reactions on time scales as short as femtoseconds under conditions that maximize the extent of reaction initiation throughout the crystal. Expand
Recent Advances in Electron Crystallography
The ultimate goal of structural biology is to understand the protein function and its physiological mechanisms by determining the three-dimensional (3D) structure. Several techniques have been usedExpand
Three-dimensional electron crystallography of protein microcrystals
TLDR
It is demonstrated that it is feasible to determine high-resolution protein structures by electron crystallography of three-dimensional crystals in an electron cryo-microscope (CryoEM) and paves the way for the implementation of a new technique, which is named ‘MicroED’, that may have wide applicability in structural biology. Expand
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 30 REFERENCES
Femtosecond X-ray protein nanocrystallography
TLDR
This work offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage, by using pulses briefer than the timescale of most damage processes. Expand
Self-terminating diffraction gates femtosecond X-ray nanocrystallography measurements.
TLDR
Measurements indicate that current X-ray free-electron laser technology should enable structural determination from submicrometre protein crystals with atomic resolution, and the shortest apparent pulse lengths occur at the highest resolution. Expand
Structure-factor analysis of femtosecond microdiffraction patterns from protein nanocrystals.
  • R. Kirian, T. White, +12 authors J. Spence
  • Materials Science, Medicine
  • Acta crystallographica. Section A, Foundations of crystallography
  • 2011
A complete set of structure factors has been extracted from hundreds of thousands of femtosecond single-shot X-ray microdiffraction patterns taken from randomly oriented nanocrystals. The method ofExpand
Experimental determination of the radiation dose limit for cryocooled protein crystals.
TLDR
The calculated dose limit of 2 x 10(7) Gy for the diffracting power of cryocooled protein crystals to drop by half has been experimentally evaluated at a third-generation synchrotron source. Expand
Radiation damage in protein serial femtosecond crystallography using an x-ray free-electron laser.
TLDR
Diffraction intensities of lysozyme nanocrystals collected at the Linac Coherent Light Source using 2 keV photons were used for structure determination by molecular replacement and analyzed for radiation damage as a function of pulse length and fluence. Expand
Femtosecond protein nanocrystallography-data analysis methods.
TLDR
The number of shots needed for a required accuracy in structure factor measurement and resolution is determined, the convergence of the Monte-Carlo integration method is investigated, and X-ray diffraction patterns for Photosystem I nanocrystals are developed. Expand
CrystFEL: a software suite for snapshot serial crystallography
In order to address the specific needs of the emerging technique of `serial femtosecond crystallography', in which structural information is obtained from small crystals illuminated by an X-rayExpand
Triclinic lysozyme at 0.65 A resolution.
TLDR
The crystal structure of triclinic hen egg-white lysozyme (HEWL) has been refined against diffraction data extending to 0.65 A resolution measured at 100 K using synchrotron radiation, and the occupancies of the water sites were refined. Expand
Potential for biomolecular imaging with femtosecond X-ray pulses
TLDR
Computer simulations are used to investigate the structural information that can be recovered from the scattering of intense femtosecond X-ray pulses by single protein molecules and small assemblies and predict that ultrashort, high-intensity X-rays from free-electron lasers that are currently under development will provide a new approach to structural determinations with X- rays. Expand
Observation of decreased radiation damage at higher dose rates in room temperature protein crystallography.
TLDR
Results of intensity-loss analysis unexpectedly showed that the dose tolerated by a crystal is dependent on the dose rate according to a positive linear relationship (99% correlation coefficient); a 60% increase in dose rate gave a 4-fold increase in crystal lifetime over the range studied. Expand
...
1
2
3
...