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Title: Towards phasing using high X-ray intensity

X-ray free-electron lasers (XFELs) show great promise for macromolecular structure determination from sub-micrometre-sized crystals, using the emerging method of serial femtosecond crystallography. The extreme brightness of the XFEL radiation can multiply ionize most, if not all, atoms in a protein, causing their scattering factors to change during the pulse, with a preferential `bleaching' of heavy atoms. This paper investigates the effects of electronic damage on experimental data collected from a Gd derivative of lysozyme microcrystals at different X-ray intensities, and the degree of ionization of Gd atoms is quantified from phased difference Fourier maps. In conclusion, a pattern sorting scheme is proposed to maximize the ionization contrast and the way in which the local electronic damage can be used for a new experimental phasing method is discussed.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [4] ;  [5] ;  [6] ;  [7] ;  [5] ;  [8] ;  [5] ;  [5] ;  [9] ;  [10] ;  [5] ;  [11]
  1. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  2. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging, Hamburg (Germany)
  3. MPI for Medical Research, Heidelberg (Germany)
  4. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  5. Max Planck Inst. for Medical Research, Heidelberg (Germany)
  6. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  7. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Uppsala Univ., Uppsala (Sweden)
  8. European Molecular Biology Lab. (EMBL), Grenoble (France)
  9. Uppsala Univ., Uppsala (Sweden)
  10. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging, Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  11. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging, Hamburg (Germany)
Publication Date:
Grant/Contract Number:
AC03-76SF00515
Type:
Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 2; Journal Issue: 6; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Research Org:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC)
Country of Publication:
United States
Language:
English
Subject:
74 ATOMIC AND MOLECULAR PHYSICS; serial femtosecond crystallography; high-intensity phasing; radiation damage; electronic damage; X-ray free-electron lasers; high XFEL doses
OSTI Identifier:
1228035

Galli, Lorenzo, Son, Sang -Kil, Barends, Thomas R. M., White, Thomas A., Barty, Anton, Botha, Sabine, Boutet, Sébastien, Caleman, Carl, Doak, R. Bruce, Nanao, Max H., Nass, Karol, Shoeman, Robert L., Timneanu, Nicusor, Santra, Robin, Schlichting, Ilme, and Chapman, Henry N.. Towards phasing using high X-ray intensity. United States: N. p., Web. doi:10.1107/S2052252515014049.
Galli, Lorenzo, Son, Sang -Kil, Barends, Thomas R. M., White, Thomas A., Barty, Anton, Botha, Sabine, Boutet, Sébastien, Caleman, Carl, Doak, R. Bruce, Nanao, Max H., Nass, Karol, Shoeman, Robert L., Timneanu, Nicusor, Santra, Robin, Schlichting, Ilme, & Chapman, Henry N.. Towards phasing using high X-ray intensity. United States. doi:10.1107/S2052252515014049.
Galli, Lorenzo, Son, Sang -Kil, Barends, Thomas R. M., White, Thomas A., Barty, Anton, Botha, Sabine, Boutet, Sébastien, Caleman, Carl, Doak, R. Bruce, Nanao, Max H., Nass, Karol, Shoeman, Robert L., Timneanu, Nicusor, Santra, Robin, Schlichting, Ilme, and Chapman, Henry N.. 2015. "Towards phasing using high X-ray intensity". United States. doi:10.1107/S2052252515014049. https://www.osti.gov/servlets/purl/1228035.
@article{osti_1228035,
title = {Towards phasing using high X-ray intensity},
author = {Galli, Lorenzo and Son, Sang -Kil and Barends, Thomas R. M. and White, Thomas A. and Barty, Anton and Botha, Sabine and Boutet, Sébastien and Caleman, Carl and Doak, R. Bruce and Nanao, Max H. and Nass, Karol and Shoeman, Robert L. and Timneanu, Nicusor and Santra, Robin and Schlichting, Ilme and Chapman, Henry N.},
abstractNote = {X-ray free-electron lasers (XFELs) show great promise for macromolecular structure determination from sub-micrometre-sized crystals, using the emerging method of serial femtosecond crystallography. The extreme brightness of the XFEL radiation can multiply ionize most, if not all, atoms in a protein, causing their scattering factors to change during the pulse, with a preferential `bleaching' of heavy atoms. This paper investigates the effects of electronic damage on experimental data collected from a Gd derivative of lysozyme microcrystals at different X-ray intensities, and the degree of ionization of Gd atoms is quantified from phased difference Fourier maps. In conclusion, a pattern sorting scheme is proposed to maximize the ionization contrast and the way in which the local electronic damage can be used for a new experimental phasing method is discussed.},
doi = {10.1107/S2052252515014049},
journal = {IUCrJ},
number = 6,
volume = 2,
place = {United States},
year = {2015},
month = {9}
}