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Drop-on-demand sample delivery for studying biocatalysts in action at X-ray free-electron lasers

DOI: 10.1038/nmeth.4195 DOI Help

Authors: Franklin D Fuller (Lawrence Berkeley National Laboratory) , Sheraz Gul (Lawrence Berkeley National Laboratory) , Ruchira Chatterjee (Lawrence Berkeley National Laboratory) , E. Sethe Burgie (Washington University in St. Louis) , Iris D. Young (Lawrence Berkeley National Laboratory) , Hugo Lebrette (Stockholm University) , Vivek Srinivas (Stockholm University) , Aaron Brewster (Lawrence Berkeley National Laboratory) , Tara Michels-clark (Lawrence Berkeley National Laboratory) , Jonathan A Clinger (Rice University, USA) , Babak Andi (National Synchrotron Light Source II) , Mohamed Ibrahim (Humboldt-Universität zu Berlin) , Ernest Pastor (Lawrence Berkeley National Laboratory) , Casper De Lichtenberg (Umeå Universitet) , Rana Hussein (Humboldt-Universität zu Berlin) , Christopher J Pollock (Pennsylvania State University) , Miao Zhang (Humboldt-Universität zu Berlin) , Claudiu A Stan (SLAC National Accelerator Laboratory) , Thomas Kroll (SLAC National Accelerator Laboratory) , Thomas Fransson (SLAC National Accelerator Laboratory) , Clemens Weninger (SLAC National Accelerator Laboratory) , Markus Kubin (Institute for Methods and Instrumentation on Synchrotron Radiation Research, Helmholtz Zentrum Berlin für Materialien und Energie) , Pierre Aller (Diamond Light Source) , Louise Lassalle (Lawrence Berkeley National Laboratory) , Philipp Braeuer (Diamond Light Source; University of Oxford) , Mitchell D. Miller (Rice University, USA) , Muhamed Amin (Lawrence Berkeley National Laboratory) , Sergey Koroidov (Umeå Universitet; SLAC National Accelerator Laboratory) , Christian G. Roessler (National Synchrotron Light Source II) , Marc Allaire (Lawrence Berkeley National Laboratory) , Raymond G Sierra (SLAC National Accelerator Laboratory) , Peter T. Docker (Diamond Light Source) , James M. Glownia (SLAC National Accelerator Laboratory) , Silke Nelson (SLAC National Accelerator Laboratory) , Jason E Koglin (SLAC National Accelerator Laboratory) , Diling Zhu (SLAC National Accelerator Laboratory) , Matthieu Chollet (SLAC National Accelerator Laboratory) , Sanghoon Song (SLAC National Accelerator Laboratory) , Henrik Lemke (SLAC National Accelerator Laboratory) , Mengning Liang (SLAC National Accelerator Laboratory) , Dimosthenis Sokaras (SLAC National Accelerator Laboratory) , Roberto Alonso-mori (SLAC National Accelerator Laboratory) , Athina Zouni (Humboldt-Universität zu Berlin) , Johannes Messinger (Umeå Universitet; Uppsala University) , Uwe Bergmann (SLAC National Accelerator Laboratory) , Amie K. Boal (Pennsylvania State University) , J. Martin Bollinger (Pennsylvania State University) , Carsten Krebs (Pennsylvania State University) , Martin Högbom (Stockholm University; Stanford University) , George N. Phillips (Rice University, USA) , Richard D. Vierstra (University in St. Louis) , Nicholas K Sauter (Lawrence Berkeley National Laboratory) , Allen M. Orville (Diamond Light Source) , Jan Kern (Lawrence Berkeley National Laboratory; SLAC National Accelerator Laboratory) , Vittal K Yachandra (Lawrence Berkeley National Laboratory) , Junko Yano (Lawrence Berkeley National Laboratory)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Methods , VOL 5

State: Published (Approved)
Published: February 2017

Abstract: X-ray crystallography at X-ray free-electron laser sources is a powerful method for studying macromolecules at biologically relevant temperatures. Moreover, when combined with complementary techniques like X-ray emission spectroscopy, both global structures and chemical properties of metalloenzymes can be obtained concurrently, providing insights into the interplay between the protein structure and dynamics and the chemistry at an active site. The implementation of such a multimodal approach can be compromised by conflicting requirements to optimize each individual method. In particular, the method used for sample delivery greatly affects the data quality. We present here a robust way of delivering controlled sample amounts on demand using acoustic droplet ejection coupled with a conveyor belt drive that is optimized for crystallography and spectroscopy measurements of photochemical and chemical reactions over a wide range of time scales. Studies with photosystem II, the phytochrome photoreceptor, and ribonucleotide reductase R2 illustrate the power and versatility of this method.

Journal Keywords: Biocatalysis; Biophysical methods; Enzymes; Molecular biophysics; Nanocrystallography

Subject Areas: Technique Development, Biology and Bio-materials, Chemistry


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