Author:
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 Božidar,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
Abstract
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.
Publisher
Springer Science and Business Media LLC