Journal Article

Membrane protein megahertz crystallography at the European XFEL

White, Thomas A., Cruz Villarreal, Jorvani, Bruce, Barry D., Kirian, Richard A., Mondal, Jyotirmoy, Mariani, Valerio, Stander, Natasha E., Shelby, Megan L., Wiedorn, Max O., Weinhausen, Britta, Jones, Alex, Bean, Richard, Zook, James D., Gevorkov, Yaroslav, Bielecki, Johan, Xu, Chen, Letrun, Romain, Mazalova, Victoria, Danilevski, Cyril, Fangohr, Hans, Zatsepin, Nadia A., Sato, Tokushi, Ketawala, Gihan K., Mancuso, Adrian P., Ourmazd, Abbas, Barty, Anton, Luna-Chavez, Cesar, Grant, Thomas D., Knoska, Juraj, Coleman, Matthew A., Januschek, Friederike, Fromme, Petra, Kuhn, Manuela, Gisriel, Chris, Schwander, Peter, Bajt, Saša, Previtali, Gianpietro, Szuba, Janusz, Hunter, Mark S., Maia, Luis, Discianno, Erin, Dörner, Katerina, Echelmeier, Austin, Boukhelef, Djelloul, Sztuk-Dambietz, Jolanta, Silenzi, Alessandro, Schmidt, Marius, Spence, John C. H., Botha, Sabine, Brockhauser, Sandor, Michelat, Thomas, Yang, Jay-How, Sarrou, Iosifina, Kim, Yoonhee, Martin-Garcia, Jose M., Abdellatif, Mohamed H., Al-Qudami, Nasser, Hosseinizadeh, Ahmad, Yefanov, Oleksandr M., Brehm, Gerrit, Manetti, Maurizio, Chapman, Henry N

2019

Overview

The world’s first superconducting megahertz repetition rate hard X-ray free-electron laser (XFEL), the European XFEL, began operation in 2017, featuring a unique pulse train structure with 886 ns between pulses. With its rapid pulse rate, the European XFEL may alleviate some of the increasing demand for XFEL beamtime, particularly for membrane protein serial femtosecond crystallography (SFX), leveraging orders-of-magnitude faster data collection. Here, we report the first membrane protein megahertz SFX experiment, where we determined a 2.9 Å-resolution SFX structure of the large membrane protein complex, Photosystem I, a > 1 MDa complex containing 36 protein subunits and 381 cofactors. We address challenges to megahertz SFX for membrane protein complexes, including growth of large quantities of crystals and the large molecular and unit cell size that influence data collection and analysis. The results imply that megahertz crystallography could have an important impact on structure determination of large protein complexes with XFELs. The European X-ray free-electron laser (EuXFEL) in Hamburg is the first XFEL with a megahertz repetition rate. Here the authors present the 2.9 Å structure of the large membrane protein complex Photosystem I from T. elongatus that was determined at the EuXFEL.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 631/45/535/1266 / 631/45/535/1266/1265 / 631/535 / 82/16 / 82/83 / Cell size / Cofactors / Crystallography / Crystals / Cyanobacteria - metabolism / Data collection / Electrons / Free electron lasers / Humanities and Social Sciences / Lasers / Membrane proteins / Membrane Proteins - chemistry / Membranes / Models, Molecular / multidisciplinary / Photosystem I / Photosystem I Protein Complex - chemistry / Photosystem I Protein Complex - isolation & purification / Protein structure / Proteins / Pulse rate / Science / Science (multidisciplinary) / Static Electricity / Synchrotrons / Thermosynechococcus / Unit cell / X-Rays