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 language | English (US) |
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Pages (from-to) | 73-78 |
Number of pages | 6 |
Journal | Nature |
Volume | 470 |
Issue number | 7332 |
DOIs | |
State | Published - Feb 3 2011 |
ASJC Scopus subject areas
- General
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Femtosecond X-ray protein Nanocrystallography
Chapman, H. N. (Contributor), Fromme, P. (Contributor), Barty, A. (Contributor), White, T. A. (Contributor), Kirian, R. (Contributor), Aquila, A. (Contributor), Hunter, M. S. (Contributor), Schulz, J. (Contributor), Deponte, D. P. (Contributor), Weierstall, U. (Contributor), Doak, R. B. (Contributor), Maia, F. R. N. C. (Contributor), Martin, A. V. (Contributor), Schlichting, I. (Contributor), Lomb, L. (Contributor), Coppola, N. (Contributor), Shoeman, R. L. (Contributor), Epp, S. W. (Contributor), Hartmann, R. (Contributor), Rolles, D. (Contributor), Rudenko, A. (Contributor), Foucar, L. (Contributor), Kimmel, N. (Contributor), Weidenspointner, G. (Contributor), Holl, P. (Contributor), Liang, M. (Contributor), Barthelmess, M. (Contributor), Caleman, C. (Contributor), Boutet, S. (Contributor), Bogan, M. J. (Contributor), Krzywinski, J. (Contributor), Bostedt, C. (Contributor), Bajt, S. (Contributor), Gumprecht, L. (Contributor), Rudek, B. (Contributor), Erk, B. (Contributor), Schmidt, C. (Contributor), Hömke, A. (Contributor), Reich, C. (Contributor), Pietschner, D. (Contributor), Ströder, L. (Contributor), Hauser, G. (Contributor), Gorke, H. (Contributor), Ullrich, J. (Contributor), Herrmann, S. (Contributor), Schaller, G. (Contributor), Schopper, F. (Contributor), Soltau, H. (Contributor), Kühnel, K. (Contributor), Messerschmidt, M. (Contributor), Bozek, J. D. (Contributor), Hau-Riege, S. P. (Contributor), Frank, M. (Contributor), Hampton, C. Y. (Contributor), Sierra, R. G. (Contributor), Starodub, D. (Contributor), Williams, G. J. (Contributor), Hajdu, J. (Contributor), Timneanu, N. (Contributor), Seibert, M. M. (Contributor), Andreasson, J. (Contributor), Rocker, A. (Contributor), Jönsson, O. (Contributor), Svenda, M. (Contributor), Stern, S. (Contributor), Nass, K. (Contributor), Andritschke, R. (Contributor), Schröter, C. (Contributor), Krasniqi, F. (Contributor), Bott, M. (Contributor), Schmidt, K. (Contributor), Wang, X. (Contributor), Grotjohann, I. (Contributor), Holton, J. M. (Contributor), Barends, T. R. M. (Contributor), Neutze, R. (Contributor), Marchesini, S. (Contributor), Fromme, R. (Contributor), Schorb, S. (Contributor), Rupp, D. (Contributor), Adolph, M. (Contributor), Gorkhover, T. (Contributor), Andersson, I. (Contributor), Hirsemann, H. (Contributor), Potdevin, G. (Contributor), Graafsma, H. (Contributor), Nilsson, B. (Contributor) & Spence, J. C. H. (Contributor), Protein Data Bank (PDB), Feb 2 2011
DOI: 10.2210/pdb3PCQ, https://www.wwpdb.org/pdb?id=pdb_00003pcq
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