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Title: Lifetimes and spatio-temporal response of protein crystals in intense X-ray microbeams

Journal Article · · IUCrJ
 [1];  [2];  [3];  [4];  [2]
  1. Cornell Univ., Ithaca, NY (United States); Rubota Corp., Portland, OR (United States)
  2. Cornell Univ., Ithaca, NY (United States)
  3. Cornell High Energy Synchrotron, Ithaca, NY (United States)
  4. Argonne National Lab. (ANL), Argonne, IL (United States)

Serial synchrotron-based crystallography using intense microfocused X-ray beams, fast-framing detectors and protein microcrystals held at 300 K promises to expand the range of accessible structural targets and to increase overall structure-pipeline throughputs. To explore the nature and consequences of X-ray radiation damage under microbeam illumination, the time-, dose- and temperature-dependent evolution of crystal diffraction have been measured with maximum dose rates of 50 MGy s–1. At all temperatures and dose rates, the integrated diffraction intensity for a fixed crystal orientation shows non-exponential decays with dose. Non-exponential decays are a consequence of non-uniform illumination and the resulting spatial evolution of diffracted intensity within the illuminated crystal volume. To quantify radiation-damage lifetimes and the damage state of diffracting crystal regions, a revised diffraction-weighted dose (DWD) is defined and it is shown that for Gaussian beams the DWD becomes nearly independent of actual dose at large doses. An apparent delayed onset of radiation damage seen in some intensity–dose curves is in fact a consequence of damage. Intensity fluctuations at high dose rates may arise from the impulsive release of gaseous damage products. Accounting for these effects, data collection at the highest dose rates increases crystal radiation lifetimes near 300 K (but not at 100 K) by a factor of ~1.5–2 compared with those observed at conventional dose rates. As a result, improved quantification and modeling of the complex spatio-temporal evolution of protein microcrystal diffraction in intense microbeams will enable more efficient data collection, and will be essential in improving the accuracy of structure factors and structural models.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Science Foundation (NSF); USDOE
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1416056
Alternate ID(s):
OSTI ID: 1434248
Journal Information:
IUCrJ, Vol. 4, Issue 6; ISSN 2052-2525
Publisher:
International Union of CrystallographyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 21 works
Citation information provided by
Web of Science

References (58)

Determination of crystallographic intensities from sparse data journal January 2015
Modeling discrete heterogeneity in X-ray diffraction data by fitting multi-conformers journal September 2009
Room-temperature serial crystallography at synchrotron X-ray sources using slowly flowing free-standing high-viscosity microstreams journal January 2015
Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams journal April 2015
Bragg coherent diffraction imaging and metrics for radiation damage in protein micro-crystallography journal January 2017
Improved reproducibility of unit-cell parameters in macromolecular cryocrystallography by limiting dehydration during crystal mounting journal July 2014
Integrated description of protein dynamics from room-temperature X-ray crystallography and NMR journal January 2014
Mitigation of X-ray damage in macromolecular crystallography by submicrometre line focusing journal July 2013
Spatial dependence and mitigation of radiation damage by a line-focus mini-beam journal November 2010
Hidden alternative structures of proline isomerase essential for catalysis journal December 2009
Radiation damage in macromolecular crystallography: what is it and why should we care? journal March 2010
Cryocooling and radiation damage in macromolecular crystallography journal December 2005
New methods for indexing multi-lattice diffraction data journal September 2014
TakeTwo: an indexing algorithm suited to still images with known crystal parameters journal July 2016
Radiation damage in protein crystallography journal September 1976
Room-temperature serial crystallography using a kinetically optimized microfluidic device for protein crystallization and on-chip X-ray diffraction journal August 2014
A beginner's guide to radiation damage journal February 2009
The R-factor gap in macromolecular crystallography: an untapped potential for insights on accurate structures journal September 2014
An assessment of the resolution limitation due to radiation-damage in X-ray diffraction microscopy journal March 2009
Cryo-cooling in macromolecular crystallography: advantages, disadvantages and optimization journal January 1999
XDS journal January 2010
Mapping the conformational landscape of a dynamic enzyme by multitemperature and XFEL crystallography journal September 2015
Flash-cooling and annealing of protein crystals journal February 2002
Heat transfer from protein crystals: implications for flash-cooling and X-ray beam heating journal March 2003
Automated electron-density sampling reveals widespread conformational polymorphism in proteins: Electron-density Sampling Detects Hidden Ensembles journal May 2010
Cryo-electron energy loss spectroscopy: observations on vitrified hydrated specimens and radiation damage journal July 1995
Radiation decay of thaumatin crystals at three X-ray energies journal March 2015
Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array journal March 2015
Growth and disorder of macromolecular crystals: insights from atomic force microscopy and X-ray diffraction studies journal November 2004
Origin and temperature dependence of radiation damage in biological samples at cryogenic temperatures journal December 2009
Fixed target matrix for femtosecond time-resolved and in situ serial micro-crystallography journal August 2015
Will reduced radiation damage occur with very small crystals? journal April 2005
Outrunning free radicals in room-temperature macromolecular crystallography journal June 2012
Exploiting fast detectors to enter a new dimension in room-temperature crystallography journal April 2014
Macromolecular cryocrystallography?methods for cooling and mounting protein crystals at cryogenic temperatures journal November 2004
The ‘fingerprint’ that X-rays can leave on structures journal March 2000
Unit-cell volume change as a metric of radiation damage in crystals of macromolecules journal November 2002
Acoustic methods for high-throughput protein crystal mounting at next-generation macromolecular crystallographic beamlines journal August 2013
Radiation damage in protein crystals is reduced with a micron-sized X-ray beam journal March 2011
Dose dependence of radiation damage for protein crystals studied at various X-ray energies journal December 2006
How does Radiation Damage in Protein Crystals Depend on X-Ray Dose? journal January 2003
Micro-crystallography comes of age journal October 2012
Observation of Decreased Radiation Damage at Higher Dose Rates in Room Temperature Protein Crystallography journal December 2007
Room-temperature macromolecular serial crystallography using synchrotron radiation journal May 2014
Reducing radiation damage in macromolecular crystals at synchrotron sources journal March 2009
Dark progression reveals slow timescales for radiation damage between T = 180 and 240 K journal August 2011
Global radiation damage: temperature dependence, time dependence and how to outrun it journal November 2012
Temperature-dependent radiation sensitivity and order of 70S ribosome crystals journal October 2014
Glass transition in thaumatin crystals revealed through temperature-dependent radiation-sensitivity measurements journal September 2010
Slow cooling and temperature-controlled protein crystallography journal December 2009
Specific chemical and structural damage to proteins produced by synchrotron radiation journal January 2000
Recent developments in CrystFEL journal March 2016
Predicting the X-ray lifetime of protein crystals journal December 2013
RADDOSE-3D : time- and space-resolved modelling of dose in macromolecular crystallography journal June 2013
Automated diffraction image analysis and spot searching for high-throughput crystal screening journal January 2006
Radiation decay of thaumatin crystals at three X-ray energies journal July 2019
Bragg coherent diffraction imaging and metrics for radiation damage in protein micro-crystallography text January 2017
An assessment of the resolution limitation due to radiation-damage in x-ray diffraction microscopy text January 2005

Cited By (12)

Megahertz non-contact luminescence decay time cryothermometry by means of ultrafast PbI2 scintillator journal March 2019
X-ray radiation damage to biological samples: recent progress journal July 2019
High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source journal April 2019
Resolving polymorphs and radiation-driven effects in microcrystals using fixed-target serial synchrotron crystallography journal November 2018
A guide to sample delivery systems for serial crystallography journal August 2019
High-speed raster-scanning synchrotron serial microcrystallography with a high-precision piezo-scanner text January 2018
Resolution and dose dependence of radiation damage in biomolecular systems journal July 2019
High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source text January 2019
Resolving polymorphs and radiation-driven effects in microcrystals using fixed-target serial synchrotron crystallography text January 2018
Megahertz non-contact luminescence decay time cryothermometry by means of ultrafast PbI2 scintillator journal March 2019
Radiation damage and dose limits in serial synchrotron crystallography at cryo- and room temperatures journal February 2020
The Influence of Photoelectron Escape in Radiation Damage Simulations of Protein Micro-Crystallography journal June 2018

Figures / Tables (5)