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Drop-on-demand sample delivery for studying biocatalysts in action at X-ray free-electron lasers
Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Berkeley, CA 94720 USA..
Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Berkeley, CA 94720 USA..
Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Berkeley, CA 94720 USA..
Washington Univ St Louis, Dept Biol, St Louis, MO USA..
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2017 (English)In: Nature Methods, ISSN 1548-7091, E-ISSN 1548-7105, Vol. 14, no 4, 443-+ p.Article in journal (Refereed) Published
Abstract [en]

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 IIII, the phytochrome photoreceptor, and ribonucleotide reductase R2 illustrate the power and versatility of this method.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP , 2017. Vol. 14, no 4, 443-+ p.
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-320026DOI: 10.1038/nmeth.4195ISI: 000397900500029PubMedID: 28250468OAI: oai:DiVA.org:uu-320026DiVA: diva2:1088602
Funder
NIH (National Institute of Health), GM110501 GM102520 GM117126 GM055302 GM113389-01 2-P41-RR012408 8P41GM103473-16 Y1GM008003 P41GM103393 P41GM111244Wellcome trustKnut and Alice Wallenberg FoundationSwedish Cancer SocietyWenner-Gren FoundationsSwedish Research Council, 2013-541 2013-5884German Research Foundation (DFG)Swedish Energy Agency, 36648-1
Available from: 2017-04-13 Created: 2017-04-13 Last updated: 2017-04-13Bibliographically approved

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