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Nat Methods. 2017 Apr;14(4):443-449. doi: 10.1038/nmeth.4195. Epub 2017 Feb 27.

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

Author information

1
Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
2
Department of Biology, Washington University in St. Louis, St. Louis, Missouri, USA.
3
Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.
4
Department of BioSciences, Rice University, Houston, Texas, USA.
5
National Synchrotron Light Source II, Brookhaven National Laboratory, Upton, New York, USA.
6
Institut für Biologie, Humboldt-Universität zu Berlin, Berlin, Germany.
7
Institutionen för Kemi, Kemiskt Biologiskt Centrum, Umeå Universitet, Umeå, Sweden.
8
Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, USA.
9
Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
10
SSRL, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
11
LCLS, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
12
Institute for Methods and Instrumentation on Synchrotron Radiation Research, Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Berlin, Germany.
13
Diamond Light Source Limited, Harwell Science and Innovation Campus, Didcot, UK.
14
Department of Biochemistry, University of Oxford, Oxford, UK.
15
Department of Chemistry-Ångström, Molecular Biomimetics, Uppsala University, Uppsala, Sweden.
16
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania, USA.
17
Department of Chemistry, Stanford University, Stanford, California, USA.
18
Department of Chemistry, Rice University, Houston, Texas, USA.

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.

PMID:
28250468
PMCID:
PMC5376230
DOI:
10.1038/nmeth.4195
[Indexed for MEDLINE]
Free PMC Article

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