Indications of radiation damage in ferredoxin microcrystals using high-intensity X-FEL beams.
Proteins that contain metal cofactors are expected to be highly radiation sensitive since the degree of X-ray absorption correlates with the presence of high-atomic-number elements and X-ray energy. To explore the effects of local damage in serial femtosecond crystallography (SFX), Clostridium ferredoxin was used as a model system. The protein contains two [4Fe-4S] clusters that serve as sensitive probes for radiation-induced electronic and structural changes. High-dose room-temperature SFX datasets were collected at the Linac Coherent Light Source of ferredoxin microcrystals. Difference electron density maps calculated from high-dose SFX and synchrotron data show peaks at the iron positions of the clusters, indicative of decrease of atomic scattering factors due to ionization. The electron density of the two [4Fe-4S] clusters differs in the FEL data, but not in the synchrotron data. Since the clusters differ in their detailed architecture, this observation is suggestive of an influence of the molecular bonding and geometry on the atomic displacement dynamics following initial photoionization. The experiments are complemented by plasma code calculations.
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Karol
K
Nass
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Lutz
L
Foucar
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Thomas R M
TR
Barends
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Elisabeth
E
Hartmann
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Sabine
S
Botha
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Robert L
RL
Shoeman
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
R Bruce
RB
Doak
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Roberto
R
Alonso-Mori
European XFEL GmbH, Albert-Einstein-Ring 19, 22761 Hamburg, Germany.
Andrew
A
Aquila
Photon Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Saša
S
Bajt
Center for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Anton
A
Barty
Center for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Richard
R
Bean
Center for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Kenneth R
KR
Beyerlein
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Maike
M
Bublitz
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Nikolaj
N
Drachmann
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Jonas
J
Gregersen
Department of Physics and Astronomy, Uppsala University, Box 516, Uppsala 75120, Sweden.
H Olof
HO
Jönsson
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Wolfgang
W
Kabsch
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Stephan
S
Kassemeyer
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Jason E
JE
Koglin
Physikalisch-Technische Bundesanstalt (PTB), Abbestrasse 2-12, 10587 Berlin, Germany.
Michael
M
Krumrey
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Daniel
D
Mattle
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Marc
M
Messerschmidt
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Poul
P
Nissen
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Linda
L
Reinhard
Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10, Aarhus 8000, Denmark.
Oleg
O
Sitsel
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Dimosthenis
D
Sokaras
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Garth J
GJ
Williams
Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA 94550, USA.
Stefan
S
Hau-Riege
Department of Physics and Astronomy, Uppsala University, Box 516, Uppsala 75120, Sweden.
Nicusor
N
Timneanu
Center for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Carl
C
Caleman
Center for Free-Electron Laser Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Henry N
HN
Chapman
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Sébastien
S
Boutet
Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Ilme
I
Schlichting
D003013:Clostridium / Q000528:radiation effects; D018360:Crystallography, X-Ray / Q000379:methods; D004307:Dose-Response Relationship, Radiation; D005288:Ferredoxins / Q000528:radiation effects*; D006801:Humans; D008667:Metalloproteins / Q000528:radiation effects*; D008958:Models, Molecular; D011832:Radiation Injuries; D012680:Sensitivity and Specificity; D017356:Synchrotrons
D005288:Ferredoxins; D008667:Metalloproteins
SFX; free-electron laser; metalloprotein; protein crystallography; radiation damage; serial femtosecond crystallography