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 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; 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 Spence; Lois Pollack; Henry N. Chapman; Saša Bajt

doi:10.1038/srep44628

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 Kirian, Anton Barty, Valerio Mariani, Aleksandra Tolstikova, Luigi Adriano, Salah Awel, Miriam Barthelmess, Katerina Dörner, P. Lourdu Xavier, Oleksandr Yefanov, Daniel R. James, Garrett NelsonDingjie 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 Spence, Lois Pollack, Henry N. Chapman, Saša Bajt

Research output: Contribution to journal › Article › peer-review

94 Scopus citations

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 language	English (US)
Article number	44628
Journal	Scientific reports
Volume	7
DOIs	https://doi.org/10.1038/srep44628
State	Published - Mar 16 2017

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10.1038/srep44628

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Oberthuer, D., Knoška, J., Wiedorn, M. O., Beyerlein, K. R., Bushnell, D. A., Kovaleva, E. G., Heymann, M., Gumprecht, L., Kirian, R., Barty, A., Mariani, V., Tolstikova, A., Adriano, L., Awel, S., Barthelmess, M., Dörner, K., Xavier, P. L., Yefanov, O., James, D. R., ... Bajt, S. (2017). Double-flow focused liquid injector for efficient serial femtosecond crystallography. Scientific reports, 7, Article 44628. https://doi.org/10.1038/srep44628

Oberthuer, D, Knoška, J, Wiedorn, MO, Beyerlein, KR, Bushnell, DA, Kovaleva, EG, Heymann, M, Gumprecht, L, Kirian, R, 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, J, 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

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title = "Double-flow focused liquid injector for efficient serial femtosecond crystallography",

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.",

author = "Dominik Oberthuer and Juraj Kno{\v s}ka and Wiedorn, {Max O.} and Beyerlein, {Kenneth R.} and Bushnell, {David A.} and Kovaleva, {Elena G.} and Michael Heymann and Lars Gumprecht and Richard Kirian and Anton Barty and Valerio Mariani and Aleksandra Tolstikova and Luigi Adriano and Salah Awel and Miriam Barthelmess and Katerina D{\"o}rner and Xavier, {P. Lourdu} and Oleksandr Yefanov and James, {Daniel R.} and Garrett Nelson and Dingjie Wang and George Calvey and Yujie Chen and Andrea Schmidt and Michael Szczepek and Stefan Frielingsdorf and Oliver Lenz and Edward Snell and Robinson, {Philip J.} and B{\"o}idar {\v S}arler and Grega Bel{\v s}ak and Marjan Ma{\v c}ek and Fabian Wilde and Andrew Aquila and S{\'e}bastien Boutet and Mengning Liang and Hunter, {Mark S.} and Patrick Scheerer and Lipscomb, {John D.} and Uwe Weierstall and Kornberg, {Roger D.} and John Spence and Lois Pollack and Chapman, {Henry N.} and Sa{\v s}a Bajt",

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year = "2017",

month = mar,

day = "16",

doi = "10.1038/srep44628",

language = "English (US)",

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T1 - Double-flow focused liquid injector for efficient serial femtosecond crystallography

AU - Oberthuer, Dominik

AU - Knoška, Juraj

AU - Wiedorn, Max O.

AU - Beyerlein, Kenneth R.

AU - Bushnell, David A.

AU - Kovaleva, Elena G.

AU - Heymann, Michael

AU - Gumprecht, Lars

AU - Kirian, Richard

AU - Barty, Anton

AU - Mariani, Valerio

AU - Tolstikova, Aleksandra

AU - Adriano, Luigi

AU - Awel, Salah

AU - Barthelmess, Miriam

AU - Dörner, Katerina

AU - Xavier, P. Lourdu

AU - Yefanov, Oleksandr

AU - James, Daniel R.

AU - Nelson, Garrett

AU - Wang, Dingjie

AU - Calvey, George

AU - Chen, Yujie

AU - Schmidt, Andrea

AU - Szczepek, Michael

AU - Frielingsdorf, Stefan

AU - Lenz, Oliver

AU - Snell, Edward

AU - Robinson, Philip J.

AU - Šarler, Böidar

AU - Belšak, Grega

AU - Maček, Marjan

AU - Wilde, Fabian

AU - Aquila, Andrew

AU - Boutet, Sébastien

AU - Liang, Mengning

AU - Hunter, Mark S.

AU - Scheerer, Patrick

AU - Lipscomb, John D.

AU - Weierstall, Uwe

AU - Kornberg, Roger D.

AU - Spence, John

AU - Pollack, Lois

AU - Chapman, Henry N.

AU - Bajt, Saša

PY - 2017/3/16

Y1 - 2017/3/16

N2 - 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.

AB - 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.

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Double-flow focused liquid injector for efficient serial femtosecond crystallography

Abstract

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12 Subunit RNA Polymerase II at Room Temperature collected using SFX

SFX structure of Cydia pomonella granulovirus using a double flow-focusing nozzle

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Double-flow focused liquid injector for efficient serial femtosecond crystallography

Abstract

ASJC Scopus subject areas

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Datasets

12 Subunit RNA Polymerase II at Room Temperature collected using SFX

SFX structure of Cydia pomonella granulovirus using a double flow-focusing nozzle

Cite this