Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Observation of a single protein by ultrafast X-ray diffraction

Journal Article · · Light, Science & Applications
 [1];  [2];  [3];  [3];  [4];  [5];  [6];  [7];  [2];  [8];  [9];  [1];  [10];  [1];  [2];  [11];  [12];  [3];  [3];  [13] more »;  [14];  [15];  [3];  [1];  [3];  [3];  [2];  [8];  [3];  [16];  [1];  [3];  [1];  [17];  [14];  [18];  [3];  [17];  [14];  [19];  [20];  [2];  [14];  [3];  [21] « less
  1. Uppsala Univ. (Sweden)
  2. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  3. European XFEL, Schenefeld (Germany)
  4. Science and Technology Facilities Council (STFC), Oxford (United Kingdom). Diamond Light Source, Ltd.
  5. University of Stavanger (Norway)
  6. Leibniz Institute for Experimental Virology (Germany)
  7. Univ. degli Studi di Milano (Italy)
  8. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science
  9. Uppsala Univ. (Sweden); Cornell Univ., Ithaca, NY (United States)
  10. PNSensor, München (Germany)
  11. European XFEL, Schenefeld (Germany); KTH Royal Inst. of Technology, Stockholm (Sweden)
  12. ELI Beamlines, Prague (Czech Republic)
  13. Centre for Structural Systems Biology (Germany); University Medical Center Hamburg-Eppendorf (UKE)
  14. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science; Univ. of Hamburg (Germany)
  15. European XFEL, Schenefeld (Germany); La Trobe Univ., Melbourne, VIC (Australia)
  16. European XFEL, Schenefeld (Germany); Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  17. Univ. of Hamburg (Germany); Max Planck Institute for the Structure and Dynamics of Matter (Germany); Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science
  18. European XFEL, Schenefeld (Germany); Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science; Max Planck Institute for the Structure and Dynamics of Matter (Germany)
  19. KTH Royal Inst. of Technology, Stockholm (Sweden)
  20. Centre for Structural Systems Biology (Germany); Univ. of Hamburg (Germany)
  21. Uppsala Univ. (Sweden); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
+ Show Author Affiliations

The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired many. It was one of the arguments for building X-ray free-electron lasers. According to theory, the extremely intense pulses provide sufficient signal to dispense with using crystals as an amplifier, and the ultrashort pulse duration permits capturing the diffraction data before the sample inevitably explodes. This was first demonstrated on biological samples a decade ago on the giant mimivirus. Since then, a large collaboration has been pushing the limit of the smallest sample that can be imaged. The ability to capture snapshots on the timescale of atomic vibrations, while keeping the sample at room temperature, may allow probing the entire conformational phase space of macromolecules. Here we show the first observation of an X-ray diffraction pattern from a single protein, that of Escherichia coli GroEL which at 14 nm in diameter is the smallest biological sample ever imaged by X-rays, and demonstrate that the concept of diffraction before destruction extends to single proteins. From the pattern, it is possible to determine the approximate orientation of the protein. Our experiment demonstrates the feasibility of ultrafast imaging of single proteins, opening the way to single-molecule time-resolved studies on the femtosecond timescale.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC)
Sponsoring Organization:
USDOE
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
2471149
Journal Information:
Light, Science & Applications, Journal Name: Light, Science & Applications Journal Issue: 1 Vol. 13; ISSN 2047-7538
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English

References (53)

UCSF Chimera?A visualization system for exploratory research and analysis journal January 2004
The Chaperonin ATPase Cycle: Mechanism of Allosteric Switching and Movements of Substrate-Binding Domains in GroEL journal October 1996
Characterising electrosprayed biomolecules using tandem-MS—the noncovalent GroEL chaperonin assembly journal August 2004
Residual counter ions can stabilise a large protein complex in the gas phase journal December 2010
Exploring the Structural Dynamics of the E.coli Chaperonin GroEL Using Translation-libration-screw Crystallographic Refinement of Intermediate States journal September 2004
Gctf: Real-time CTF determination and correction journal January 2016
Time-resolved cryo-electron microscopy: Recent progress journal December 2017
Large-format, high-speed, X-ray pnCCDs combined with electron and ion imaging spectrometers in a multipurpose chamber for experiments at 4th generation light sources
  • Strüder, Lothar; Epp, Sascha; Rolles, Daniel
  • Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, Vol. 614, Issue 3 https://doi.org/10.1016/j.nima.2009.12.053
journal March 2010
GROMACS: High performance molecular simulations through multi-level parallelism from laptops to supercomputers journal September 2015
Seeing GroEL at 6 Å Resolution by Single Particle Electron Cryomicroscopy journal July 2004
The GroEL–GroES Chaperonin Machine: A Nano-Cage for Protein Folding journal January 2016
Improving the Performance of a Quadrupole Time-of-Flight Instrument for Macromolecular Mass Spectrometry journal November 2006
Detection of the Intact GroEL Chaperonin Assembly by Mass Spectrometry journal April 1999
Tandem Differential Mobility Analysis-Mass Spectrometry Reveals Partial Gas-Phase Collapse of the GroEL Complex journal April 2011
Potential for biomolecular imaging with femtosecond X-ray pulses journal August 2000
The crystal structure of the bacterial chaperonln GroEL at 2.8 Å journal October 1994
Single mimivirus particles intercepted and imaged with an X-ray laser journal February 2011
Femtosecond X-ray protein nanocrystallography journal February 2011
Nonlinear lattice dynamics as a basis for enhanced superconductivity in YBa2Cu3O6.5 journal December 2014
Imaging single cells in a beam of live cyanobacteria with an X-ray laser journal February 2015
The Coherent X-ray Imaging Data Bank journal August 2012
High-sensitivity Orbitrap mass analysis of intact macromolecular assemblies journal October 2012
The development of cryo-EM into a mainstream structural biology technique journal December 2015
cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination journal February 2017
MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy journal February 2017
Femtosecond diffractive imaging with a soft-X-ray free-electron laser journal November 2006
A MHz-repetition-rate hard X-ray free-electron laser driven by a superconducting linear accelerator journal May 2020
SciPy 1.0: fundamental algorithms for scientific computing in Python journal February 2020
SPHIRE-crYOLO is a fast and accurate fully automated particle picker for cryo-EM journal June 2019
Structural stability of electrosprayed proteins: temperature and hydration effects journal January 2009
The linac coherent light source single particle imaging road map journal July 2015
Subunit conformational variation within individual GroEL oligomers resolved by Cryo-EM journal July 2017
Design and development of electronics for the EuXFEL clock and control system journal January 2012
Reconstruction algorithm for single-particle diffraction imaging experiments journal August 2009
Structural variability and the incoherent addition of scattered intensities in single-particle diffraction journal September 2009
Sacrificial Tamper Slows Down Sample Explosion in FLASH Diffraction Experiments journal February 2010
Three-Dimensional Reconstruction of the Giant Mimivirus Particle with an X-Ray Free-Electron Laser journal March 2015
Hummingbird : monitoring and analyzing flash X-ray imaging experiments in real time journal April 2016
Condor : a simulation tool for flash X-ray imaging journal July 2016
Post-sample aperture for low background diffraction experiments at X-ray free-electron lasers journal October 2017
Operation of X-ray gas monitors at the European XFEL journal June 2019
The 1-Megapixel pnCCD detector for the Small Quantum Systems Instrument at the European XFEL: system and operation aspects journal January 2021
Experimental strategies for imaging bioparticles with femtosecond hard X-ray pulses journal April 2017
Rayleigh-scattering microscopy for tracking and sizing nanoparticles in focused aerosol beams journal September 2018
The MiniSDD-Based 1-Mpixel Camera of the DSSC Project for the European XFEL journal June 2021
Electrospray sample injection for single-particle imaging with x-ray lasers journal May 2019
Time-resolved serial crystallography captures high-resolution intermediates of photoactive yellow protein journal December 2014
Observation of the fastest chemical processes in the radiolysis of water journal January 2020
Experimental strategies for imaging bioparticles with femtosecond hard X-ray pulses
  • Daurer, Benedikt
  • Coherent X-ray Imaging Data Bank (Lawrence Berkeley National Laboratory); University of Gothenburg, University of Jyvaskyla, University of Helsinki, CFEL, The University of Tokyo, Arizona State University, La Trobe University, University of Wisconsin, SLAC National Accelerator Laboratory, RIKEN SPring-8 Center, University of Chicago, Argonne National Laboratory, Northeastern Illinois University https://doi.org/10.11577/1349716
dataset January 2016
Low-signal limit of X-ray single particle diffractive imaging journal January 2019
Overview of Acquisition and Control Electronics and Concepts for Experiments and Beam Transport at the European XFEL text January 2020
Photon Beam Transport and Scientific Instruments at the European XFEL journal June 2017
New tools for automated high-resolution cryo-EM structure determination in RELION-3 journal November 2018

Cited By (1)

Observation of a single protein by ultrafast X-ray diffraction dataset January 2024

Similar Records

Femtosecond Diffractive Imaging with a Soft-X-ray Free-Electron Laser
Journal Article · Sun Mar 12 23:00:00 EST 2006 · Nature Physics, vol. 2, no. 12, November 12, 2006, pp. 839-843 · OSTI ID:900470
Femtosecond Diffractive Imaging with a Soft-X-Ray Free-Electron Laser
Journal Article · Thu Oct 07 00:00:00 EDT 2010 · Nature PHys.2:839, 2006 · OSTI ID:992877
Challenges and opportunities in attosecond and XFEL science
Journal Article · Mon Jan 28 23:00:00 EST 2019 · Nature Reviews Physics · OSTI ID:1514996

Related Subjects

71 CLASSICAL AND QUANTUM MECHANICS
GENERAL PHYSICS
imaging
sensing
x-rays