Femtosecond Three-Dimensional Imaging of Single-Protein with Hard X-ray Laser
Paul Lourdu Xavier, Andrew J. Morgan, Johan Bielecki, Amit K. Samanta, Simon Dold, Chufeng Li, Xinyue Gao, Spencer K. Passmore, Ruslan P. Kurta, Tim B. Berberich, Emiliano De Santis, Wenhui Zhang, Juncheng E, Faisal H. M. Koua, Romain Letrun, Luca Gelisio, Oleksii Turkot, Egor Sobolev, Lukas V. Haas, Jannik Lübke, Jingxuan He, Madeline B. F. Memovich, Kevin Janson, Stefanie Lenzen, Safi Rafie-Zinedine, Mansi Butola, Oleksandr Yefanov, Fabian Trost, Chan Kim, Tomas Popelar, Tokushi Sato, Yevheniy Ovcharenko, Rebecca Boll, Sergey Usenko, Thomas M. Baumann, Alberto De Fanis, Björn Senftleben, Matteo Porro, Andrea Castoldi, Ibrahym Dourki, Cyril Danilevski, Ekaterina Round, Christina Schmidt, Huijong Han, Roberto Alvarez, Erin C. Yang, Shunzhi Wang, Marcos C. Miranda, Hao Shen, Alexis Courbet, Nathan M. Ennist, Ervin Chia, Cornelia Cazey, Carolin Seuring, Ruojie Sha, Maia Azubel, David A. Bushnell, Sébastien Boutet, Sergey Ovchinnikov, Adrian P. Mancuso, Michael Meyer, Richard Bean, Anton Barty, Filipe R. N. C. Maia, Richard A. Kirian, Thomas D. Grant, N. Duane Loh, Carl Caleman, Erik G. Marklund, Nadia Zatsepin, Andrew V. Martin, Saša Bajt, Roger D. Kornberg, David Baker, Jochen Küpper, Henry N. Chapman
Abstract
The extremely intense pulses of X-ray free-electron lasers (XFELs) have enabled imaging of radiation-sensitive samples, such as macromolecular microcrystals, beyond radiation damage limits. These sources have the potential to deliver biomolecular single-particle imaging, similar to cryo-electron microscopy but without the need for cryo-fixation and with temporal resolution from femtoseconds to milliseconds. While this possibility was recognized before XFELs were built, the biological single-particle imaging work-flow has previously only been demonstrated on large virus particles. Based on decades of improvements in X-ray beam focusing, particle delivery, diffraction detection, and advanced analysis, here we demonstrate imaging of a single molecular complex, the giant-hemoglobin erythrocruorin (Ery) with X-ray laser pulses. Two-dimensional classes of diffraction patterns could be reconstructed to 15 Angstrom resolution, and 3D images to approximately 20 Angstrom, while the 3D merged intensity in reciprocal space extended beyond 20 Angstrom. The resolution discrepancy is likely due to heterogeneity caused by gas-phase compaction of the complexes. With increased throughput, this approach could be used to reveal in-situ structural details during mass spectrometry studies of biomolecules, while improvements in sample delivery may provide ultrafast snapshot imaging of biological single-particles in their native-state beyond the limitations of radiation damage.
Create a lesson
Related papers
Efficient Transport Networks through Edge Pruning by Relative Resistance
Mathieu Le Verge-Serandour, Viktor Vasilev, Lara Szeimies et al.
Boundary-contact repolarization provides a mechanism for directional guidance of neural crest cell collectives
Aditya Shankar Paspunurwar, Hector Gomez
Three-dimensional imaging of isolated membrane-protein complexes in vacuo with an X-ray laser
Kartik Ayyer, Tej Varma Yenupuri, Johan Bielecki et al.
Polarization of the Drosophila oocyte by active cortical exchange
Mohammadhossein Firouznia, Wen Lu, Margot Lakonishok et al.
Inverse-Designed Anisotropic Intrastromal Corneal Exoskeletons for Keratoconus: A Three-Dimensional Computational Feasibility Study
J. Sumaya-Martinez, Alan Altamirano
Spatiotemporal Oscillations Driven by Spatial Heterogeneity and Growth
Lewis S. Mosby, Mohit P. Dalwadi, Zena Hadjivasiliou