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

Abstract

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 notmore » 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.« less

Authors:
 [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)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Science Foundation (NSF)
OSTI Identifier:
1434248
Grant/Contract Number:
AC02-06CH11357
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 4; Journal Issue: 6; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS; X-ray crystallography; intense X-ray microbeams; microcrystallography; protein crystallography; protein structure; radiation damage; serial crystallography; structural biology; structure determination

Citation Formats

Warkentin, Matthew A., Atakisi, Hakan, Hopkins, Jesse B., Walko, Donald, and Thorne, Robert E.. Lifetimes and spatio-temporal response of protein crystals in intense X-ray microbeams. United States: N. p., 2017. Web. doi:10.1107/s2052252517013495.
Warkentin, Matthew A., Atakisi, Hakan, Hopkins, Jesse B., Walko, Donald, & Thorne, Robert E.. Lifetimes and spatio-temporal response of protein crystals in intense X-ray microbeams. United States. doi:10.1107/s2052252517013495.
Warkentin, Matthew A., Atakisi, Hakan, Hopkins, Jesse B., Walko, Donald, and Thorne, Robert E.. Fri . "Lifetimes and spatio-temporal response of protein crystals in intense X-ray microbeams". United States. doi:10.1107/s2052252517013495. https://www.osti.gov/servlets/purl/1434248.
@article{osti_1434248,
title = {Lifetimes and spatio-temporal response of protein crystals in intense X-ray microbeams},
author = {Warkentin, Matthew A. and Atakisi, Hakan and Hopkins, Jesse B. and Walko, Donald and Thorne, Robert E.},
abstractNote = {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.},
doi = {10.1107/s2052252517013495},
journal = {IUCrJ},
number = 6,
volume = 4,
place = {United States},
year = {Fri Oct 13 00:00:00 EDT 2017},
month = {Fri Oct 13 00:00:00 EDT 2017}
}

Journal Article:
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