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

Franklin D. Fuller, Sheraz Gul, Ruchira Chatterjee, E. Sethe Burgie, Iris D. Young, Hugo Lebrette, Vivek Srinivas, Aaron S. Brewster, Tara Michels-Clark, Jonathan A. Clinger, Babak Andi, Mohamed Ibrahim, Ernest Pastor, Casper De Lichtenberg, Rana Hussein, Christopher J. Pollock, Miao Zhang, Claudiu A. Stan, Thomas Kroll, Thomas FranssonClemens Weninger, Markus Kubin, Pierre Aller, Louise Lassalle, Philipp Bräuer, Mitchell D. Miller, Muhamed Amin, Sergey Koroidov, Christian G. Roessler, Marc Allaire, Raymond G. Sierra, Peter T. Docker, James M. Glownia, Silke Nelson, Jason E. Koglin, Diling Zhu, Matthieu Chollet, Sanghoon Song, Henrik Lemke, Mengning Liang, Dimosthenis Sokaras, Roberto Alonso-Mori, Athina Zouni, Johannes Messinger, Uwe Bergmann, Amie K. Boal, J. Martin Bollinger, Carsten Krebs, Martin Högbom, George N. Phillips, Richard D. Vierstra, Nicholas K. Sauter, Allen M. Orville, Jan Kern, Vittal K. Yachandra, Junko Yano

Research output: Contribution to journalArticle

56 Scopus citations

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 con?icting 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.

Original languageEnglish (US)
Pages (from-to)443-449
Number of pages7
JournalNature methods
Volume14
Issue number4
DOIs
StatePublished - Feb 27 2017

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Fuller, F. D., Gul, S., Chatterjee, R., Sethe Burgie, E., Young, I. D., Lebrette, H., Srinivas, V., Brewster, A. S., Michels-Clark, T., Clinger, J. A., Andi, B., Ibrahim, M., Pastor, E., De Lichtenberg, C., Hussein, R., Pollock, C. J., Zhang, M., Stan, C. A., Kroll, T., ... Yano, J. (2017). Drop-on-demand sample delivery for studying biocatalysts in action at X-ray free-electron lasers. Nature methods, 14(4), 443-449. https://doi.org/10.1038/nmeth.4195