Femtosecond X-ray protein nanocrystallography

Henry N. Chapman, Petra Fromme, Anton Barty, Thomas A. White, Richard A. Kirian, Andrew Aquila, Mark S. Hunter, Joachim Schulz, Daniel P. Deponte, Uwe Weierstall, R. Bruce Doak, Filipe R N C Maia, Andrew V. Martin, Ilme Schlichting, Lukas Lomb, Nicola Coppola, Robert L. Shoeman, Sascha W. Epp, Robert Hartmann, Daniel RollesArtem Rudenko, Lutz Foucar, Nils Kimmel, Georg Weidenspointner, Peter Holl, Mengning Liang, Miriam Barthelmess, Carl Caleman, Sébastien Boutet, Michael J. Bogan, Jacek Krzywinski, Christoph Bostedt, Sǎa Bajt, Lars Gumprecht, Benedikt Rudek, Benjamin Erk, Carlo Schmidt, André Hömke, Christian Reich, Daniel Pietschner, Lothar Ströder, Günter Hauser, Hubert Gorke, Joachim Ullrich, Sven Herrmann, Gerhard Schaller, Florian Schopper, Heike Soltau, Kai Uwe Kühnel, Marc Messerschmidt, John D. Bozek, Stefan P. Hau-Riege, Matthias Frank, Christina Y. Hampton, Raymond G. Sierra, Dmitri Starodub, Garth J. Williams, Janos Hajdu, Nicusor Timneanu, M. Marvin Seibert, Jakob Andreasson, Andrea Rocker, Olof Jönsson, Martin Svenda, Stephan Stern, Karol Nass, Robert Andritschke, Claus Dieter Schröter, Faton Krasniqi, Mario Bott, Kevin E. Schmidt, Xiaoyu Wang, Ingo Grotjohann, James M. Holton, Thomas R M Barends, Richard Neutze, Stefano Marchesini, Raimund Fromme, Sebastian Schorb, Daniela Rupp, Marcus Adolph, Tais Gorkhover, Inger Andersson, Helmut Hirsemann, Guillaume Potdevin, Heinz Graafsma, Björn Nilsson, John C H Spence

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1209 Citations (Scopus)

Abstract

X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction snapshotsg are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (4200nm to 21/4m in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.

Original languageEnglish
Pages (from-to)73-78
Number of pages6
JournalNature
Volume470
Issue number7332
DOIs
Publication statusPublished - Feb 3 2011

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Chapman, H. N., Fromme, P., Barty, A., White, T. A., Kirian, R. A., Aquila, A., ... Spence, J. C. H. (2011). Femtosecond X-ray protein nanocrystallography. Nature, 470(7332), 73-78. https://doi.org/10.1038/nature09750