TY - JOUR
T1 - Room temperature femtosecond X-ray diffraction of photosystem II microcrystals
AU - Kern, Jan
AU - Alonso-Mori, Roberto
AU - Hellmich, Julia
AU - Tran, Rosalie
AU - Hattne, Johan
AU - Laksmono, Hartawan
AU - Glöckner, Carina
AU - Echols, Nathaniel
AU - Sierra, Raymond G.
AU - Sellberg, Jonas
AU - Lassalle-Kaiser, Benedikt
AU - Gildea, Richard J.
AU - Glatzel, Pieter
AU - Grosse-Kunstleve, Ralf W.
AU - Latimer, Matthew J.
AU - McQueen, Trevor A.
AU - DiFiore, Dörte
AU - Fry, Alan R.
AU - Messerschmidt, Marc
AU - Miahnahri, Alan
AU - Schafer, Donald W.
AU - Seibert, M. Marvin
AU - Sokaras, Dimosthenis
AU - Weng, Tsu Chien
AU - Zwart, Petrus H.
AU - White, William E.
AU - Adams, Paul D.
AU - Bogan, Michael J.
AU - Boutet, Sébastien
AU - Williams, Garth J.
AU - Messinger, Johannes
AU - Sauter, Nicholas K.
AU - Zouni, Athina
AU - Bergmann, Uwe
AU - Yano, Junko
AU - Yachandra, Vittal K.
PY - 2012/6/19
Y1 - 2012/6/19
N2 - Most of the dioxygen on earth is generated by the oxidation of water by photosystem II (PS II) using light from the sun. This light-driven, four-photon reaction is catalyzed by the Mn 4CaO 5 cluster located at the lumenal side of PS II. Various X-ray studies have been carried out at cryogenic temperatures to understand the intermediate steps involved in the water oxidation mechanism. However, the necessity for collecting data at room temperature, especially for studying the transient steps during the O-O bond formation, requires the development of new methodologies. In this paper we report room temperature X-ray diffraction data of PS II microcrystals obtained using ultrashort (<50 fs) 9 keV X-ray pulses from a hard X-ray free electron laser, namely the Linac Coherent Light Source. The results presented here demonstrate that the "probe before destroy" approach using an X-ray free electron laser works even for the highly-sensitive Mn 4CaO 5 cluster in PS II at room temperature. We show that these data are comparable to those obtained in synchrotron radiation studies as seen by the similarities in the overall structure of the helices, the protein subunits and the location of the various cofactors. This work is, therefore, an important step toward future studies for resolving the structure of the Mn 4CaO 5 cluster without any damage at room temperature, and of the reaction intermediates of PS II during O-O bond formation.
AB - Most of the dioxygen on earth is generated by the oxidation of water by photosystem II (PS II) using light from the sun. This light-driven, four-photon reaction is catalyzed by the Mn 4CaO 5 cluster located at the lumenal side of PS II. Various X-ray studies have been carried out at cryogenic temperatures to understand the intermediate steps involved in the water oxidation mechanism. However, the necessity for collecting data at room temperature, especially for studying the transient steps during the O-O bond formation, requires the development of new methodologies. In this paper we report room temperature X-ray diffraction data of PS II microcrystals obtained using ultrashort (<50 fs) 9 keV X-ray pulses from a hard X-ray free electron laser, namely the Linac Coherent Light Source. The results presented here demonstrate that the "probe before destroy" approach using an X-ray free electron laser works even for the highly-sensitive Mn 4CaO 5 cluster in PS II at room temperature. We show that these data are comparable to those obtained in synchrotron radiation studies as seen by the similarities in the overall structure of the helices, the protein subunits and the location of the various cofactors. This work is, therefore, an important step toward future studies for resolving the structure of the Mn 4CaO 5 cluster without any damage at room temperature, and of the reaction intermediates of PS II during O-O bond formation.
KW - Manganese
KW - Oxygen-evolving complex
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U2 - 10.1073/pnas.1204598109
DO - 10.1073/pnas.1204598109
M3 - Article
C2 - 22665786
AN - SCOPUS:84862545637
SN - 0027-8424
VL - 109
SP - 9721
EP - 9726
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 25
ER -