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

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

in 631/1647/2204 / 631/1647/2258/1266/1265 / 631/45/603 / 631/45/607 / 631/57/2272 / Acoustic coupling / Acoustics / Analysis / Atmospheric chemistry / BASIC BIOLOGICAL SCIENCES / Belt conveyors / Biocatalysis / Biocatalysts / Biochemistry / Bioinformatics / biokemi / Biological Microscopy / Biological Techniques / Biomedical Engineering/Biotechnology / Biophysical methods / Chemical properties / Chemical reactions / Conveyors / Crystallography / Crystallography, X-Ray - methods / Ejection / Emission spectroscopy / Enzymes / Free electron lasers / Lasers / Life Sciences / Macromolecules / Molecular biophysics / Nanocrystallography / Organic chemistry / Photochemical reactions / Photochemicals / Photosystem II / Photosystem II Protein Complex - chemistry / Phytochrome - chemistry / Protein structure / Proteins / Proteomics / Reductase / Ribonucleotide Reductases - chemistry / Robust control / Spectrometry, X-Ray Emission - methods / Spectroscopy / X-ray crystallography

2017

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

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Journal Article

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

Zouni, Athina,

Högbom, Martin,

Aller, Pierre,

Chollet, Matthieu,

Sokaras, Dimosthenis,

Sierra, Raymond G,

Weninger, Clemens,

Fransson, Thomas,

Phillips, George N,

Clinger, Jonathan A,

Alonso-Mori, Roberto,

Bergmann, Uwe,

Kern, Jan,

Messinger, Johannes,

Allaire, Marc,

Liang, Mengning,

Bräuer, Philipp,

Boal, Amie K,

Zhang, Miao,

Kroll, Thomas,

Vierstra, Richard D,

Lebrette, Hugo,

Lassalle, Louise,

Nelson, Silke,

Stan, Claudiu A,

Yachandra, Vittal K,

Brewster, Aaron S,

Andi, Babak,

Young, Iris D,

Orville, Allen M,

Song, Sanghoon,

Koglin, Jason E,

Srinivas, Vivek,

Pastor, Ernest,

de Lichtenberg, Casper,

Gul, Sheraz,

Pollock, Christopher J,

Amin, Muhamed,

Yano, Junko,

Hussein, Rana,

Bollinger, J Martin,

Lemke, Henrik,

Ibrahim, Mohamed,

Kubin, Markus,

Chatterjee, Ruchira,

Fuller, Franklin D,

Zhu, Diling,

Burgie, E Sethe,

Roessler, Christian G,

Koroidov, Sergey,

Docker, Peter T,

Sauter, Nicholas K,

Michels-Clark, Tara,

Miller, Mitchell D,

Krebs, Carsten,

Glownia, James M

2017

Overview

A robust acoustic droplet ejection–drop-on-tape method delivers samples to an X-ray free-electron laser source for combined serial femtosecond crystallography and X-ray emission spectroscopy analysis, providing detailed insights into macromolecular reaction dynamics. 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.

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Publisher

Nature Publishing Group US,Nature Publishing Group

Subject

631/1647/2204

/ 631/1647/2258/1266/1265