Double-flow focused liquid injector for efficient serial femtosecond crystallography

Dominik Oberthuer, Juraj Knoška, Max O. Wiedorn, Kenneth R. Beyerlein, David A. Bushnell, Elena G. Kovaleva, Michael Heymann, Lars Gumprecht, Richard A. Kirian, Anton Barty, Valerio Mariani, Aleksandra Tolstikova, Luigi Adriano, Salah Awel, Miriam Barthelmess, Katerina Dörner, P. Lourdu Xavier, Oleksandr Yefanov, Daniel R. James, Garrett Nelson & 25 others Dingjie Wang, George Calvey, Yujie Chen, Andrea Schmidt, Michael Szczepek, Stefan Frielingsdorf, Oliver Lenz, Edward Snell, Philip J. Robinson, Böidar Šarler, Grega Belšak, Marjan Maček, Fabian Wilde, Andrew Aquila, Sébastien Boutet, Mengning Liang, Mark S. Hunter, Patrick Scheerer, John D. Lipscomb, Uwe Weierstall, Roger D. Kornberg, John C.H. Spence, Lois Pollack, Henry N. Chapman, Saša Bajt

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam of an X-ray free-electron laser over the course of an experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability over previous generations of nozzles. We demonstrate its use to determine the first room-temperature structure of RNA polymerase II at high resolution, revealing new structural details. Moreover, the double flow-focusing nozzles were successfully tested with three other protein samples and the first room temperature structure of an extradiol ring-cleaving dioxygenase was solved by utilizing the improved operation and characteristics of these devices.

Original languageEnglish (US)
Article number44628
JournalScientific reports
Volume7
DOIs
StatePublished - Mar 16 2017

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Crystallography
Dioxygenases
Temperature
RNA Polymerase II
Lasers
X-Rays
Electrons
Equipment and Supplies
Proteins

Cite this

Oberthuer, D., Knoška, J., Wiedorn, M. O., Beyerlein, K. R., Bushnell, D. A., Kovaleva, E. G., ... Bajt, S. (2017). Double-flow focused liquid injector for efficient serial femtosecond crystallography. Scientific reports, 7, [44628]. https://doi.org/10.1038/srep44628

Double-flow focused liquid injector for efficient serial femtosecond crystallography. / Oberthuer, Dominik; Knoška, Juraj; Wiedorn, Max O.; Beyerlein, Kenneth R.; Bushnell, David A.; Kovaleva, Elena G.; Heymann, Michael; Gumprecht, Lars; Kirian, Richard A.; Barty, Anton; Mariani, Valerio; Tolstikova, Aleksandra; Adriano, Luigi; Awel, Salah; Barthelmess, Miriam; Dörner, Katerina; Xavier, P. Lourdu; Yefanov, Oleksandr; James, Daniel R.; Nelson, Garrett; Wang, Dingjie; Calvey, George; Chen, Yujie; Schmidt, Andrea; Szczepek, Michael; Frielingsdorf, Stefan; Lenz, Oliver; Snell, Edward; Robinson, Philip J.; Šarler, Böidar; Belšak, Grega; Maček, Marjan; Wilde, Fabian; Aquila, Andrew; Boutet, Sébastien; Liang, Mengning; Hunter, Mark S.; Scheerer, Patrick; Lipscomb, John D.; Weierstall, Uwe; Kornberg, Roger D.; Spence, John C.H.; Pollack, Lois; Chapman, Henry N.; Bajt, Saša.

In: Scientific reports, Vol. 7, 44628, 16.03.2017.

Research output: Contribution to journalArticle

Oberthuer, D, Knoška, J, Wiedorn, MO, Beyerlein, KR, Bushnell, DA, Kovaleva, EG, Heymann, M, Gumprecht, L, Kirian, RA, Barty, A, Mariani, V, Tolstikova, A, Adriano, L, Awel, S, Barthelmess, M, Dörner, K, Xavier, PL, Yefanov, O, James, DR, Nelson, G, Wang, D, Calvey, G, Chen, Y, Schmidt, A, Szczepek, M, Frielingsdorf, S, Lenz, O, Snell, E, Robinson, PJ, Šarler, B, Belšak, G, Maček, M, Wilde, F, Aquila, A, Boutet, S, Liang, M, Hunter, MS, Scheerer, P, Lipscomb, JD, Weierstall, U, Kornberg, RD, Spence, JCH, Pollack, L, Chapman, HN & Bajt, S 2017, 'Double-flow focused liquid injector for efficient serial femtosecond crystallography' Scientific reports, vol. 7, 44628. https://doi.org/10.1038/srep44628
Oberthuer D, Knoška J, Wiedorn MO, Beyerlein KR, Bushnell DA, Kovaleva EG et al. Double-flow focused liquid injector for efficient serial femtosecond crystallography. Scientific reports. 2017 Mar 16;7. 44628. https://doi.org/10.1038/srep44628
Oberthuer, Dominik ; Knoška, Juraj ; Wiedorn, Max O. ; Beyerlein, Kenneth R. ; Bushnell, David A. ; Kovaleva, Elena G. ; Heymann, Michael ; Gumprecht, Lars ; Kirian, Richard A. ; Barty, Anton ; Mariani, Valerio ; Tolstikova, Aleksandra ; Adriano, Luigi ; Awel, Salah ; Barthelmess, Miriam ; Dörner, Katerina ; Xavier, P. Lourdu ; Yefanov, Oleksandr ; James, Daniel R. ; Nelson, Garrett ; Wang, Dingjie ; Calvey, George ; Chen, Yujie ; Schmidt, Andrea ; Szczepek, Michael ; Frielingsdorf, Stefan ; Lenz, Oliver ; Snell, Edward ; Robinson, Philip J. ; Šarler, Böidar ; Belšak, Grega ; Maček, Marjan ; Wilde, Fabian ; Aquila, Andrew ; Boutet, Sébastien ; Liang, Mengning ; Hunter, Mark S. ; Scheerer, Patrick ; Lipscomb, John D. ; Weierstall, Uwe ; Kornberg, Roger D. ; Spence, John C.H. ; Pollack, Lois ; Chapman, Henry N. ; Bajt, Saša. / Double-flow focused liquid injector for efficient serial femtosecond crystallography. In: Scientific reports. 2017 ; Vol. 7.
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AU - Szczepek, Michael

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AU - Aquila, Andrew

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