Mix-and-diffuse serial synchrotron crystallography
Abstract
Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. Here, the success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources.
- Authors:
-
more »
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
- California State Univ., Northridge, CA (United States)
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Norwegian Univ. of Science and Technology, Trondheim (Norway)
- Uppsala Univ., Uppsala (Sweden)
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); European X-ray Free-Electron Laser Facility GmbH (XFEL), Schenefeld (Germany)
- Univ. of Oxford, Oxford (England)
- SLAC National Accelerator Lab., Menlo Park, CA (United States)
- Fachhochschule Lubeck, Lubeck (Germany)
- Publication Date:
- Research Org.:
- SLAC National Accelerator Lab., Menlo Park, CA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1410554
- Grant/Contract Number:
- AC02-76SF00515; FP7/2007-2013; ERC-614507-Küpper; DFG-EXC1074
- Resource Type:
- Accepted Manuscript
- Journal Name:
- IUCrJ
- Additional Journal Information:
- Journal Volume: 4; Journal Issue: 6; Journal ID: ISSN 2052-2525
- Publisher:
- International Union of Crystallography
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES; drug discovery; protein structure; X-ray crystallography; serial crystallography; time-resolved studies; lysozyme
Citation Formats
Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik. Mix-and-diffuse serial synchrotron crystallography. United States: N. p., 2017.
Web. doi:10.1107/S2052252517013124.
Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, & Oberthuer, Dominik. Mix-and-diffuse serial synchrotron crystallography. United States. https://doi.org/10.1107/S2052252517013124
Beyerlein, Kenneth R., Dierksmeyer, Dennis, Mariani, Valerio, Kuhn, Manuela, Sarrou, Iosifina, Ottaviano, Angelica, Awel, Salah, Knoska, Juraj, Fuglerud, Silje, Jönsson, Olof, Stern, Stephan, Wiedorn, Max O., Yefanov, Oleksandr, Adriano, Luigi, Bean, Richard, Burkhardt, Anja, Fischer, Pontus, Heymann, Michael, Horke, Daniel A., Jungnickel, Katharina E. J., Kovaleva, Elena, Lorbeer, Olga, Metz, Markus, Meyer, Jan, Morgan, Andrew, Pande, Kanupriya, Panneerselvam, Saravanan, Seuring, Carolin, Tolstikova, Aleksandra, Lieske, Julia, Aplin, Steve, Roessle, Manfred, White, Thomas A., Chapman, Henry N., Meents, Alke, and Oberthuer, Dominik. Mon .
"Mix-and-diffuse serial synchrotron crystallography". United States. https://doi.org/10.1107/S2052252517013124. https://www.osti.gov/servlets/purl/1410554.
@article{osti_1410554,
title = {Mix-and-diffuse serial synchrotron crystallography},
author = {Beyerlein, Kenneth R. and Dierksmeyer, Dennis and Mariani, Valerio and Kuhn, Manuela and Sarrou, Iosifina and Ottaviano, Angelica and Awel, Salah and Knoska, Juraj and Fuglerud, Silje and Jönsson, Olof and Stern, Stephan and Wiedorn, Max O. and Yefanov, Oleksandr and Adriano, Luigi and Bean, Richard and Burkhardt, Anja and Fischer, Pontus and Heymann, Michael and Horke, Daniel A. and Jungnickel, Katharina E. J. and Kovaleva, Elena and Lorbeer, Olga and Metz, Markus and Meyer, Jan and Morgan, Andrew and Pande, Kanupriya and Panneerselvam, Saravanan and Seuring, Carolin and Tolstikova, Aleksandra and Lieske, Julia and Aplin, Steve and Roessle, Manfred and White, Thomas A. and Chapman, Henry N. and Meents, Alke and Oberthuer, Dominik},
abstractNote = {Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. Here, the success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources.},
doi = {10.1107/S2052252517013124},
journal = {IUCrJ},
number = 6,
volume = 4,
place = {United States},
year = {2017},
month = {10}
}
Web of Science
Figures / Tables:

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Strategies for sample delivery for femtosecond crystallography
text, January 2019
- Martiel, Isabelle; Müller-Werkmeister, Henrike M.; Cohen, Aina E.
- International Union of Crystallography (IUCr)
Rapid sample delivery for megahertz serial crystallography at X-ray FELs
text, January 2018
- Wiedorn, Max Oliver; Awel, Salah; Morgan, Andrew J.
- TUHH Universitätsbibliothek
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies
text, January 2019
- Lieske, Julia; Cerv, Maximilian; Kreida, Stefan
- Deutsches Elektronen-Synchrotron, DESY, Hamburg
High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source
text, January 2019
- Martin-Garcia, Jose M.; Zhu, Lan; Mendez, Derek
- International Union of Crystallography (IUCr)
Serial protein crystallography in an electron microscope
text, January 2020
- Bücker, Robert; Hogan-Lamarre, Pascal; Mehrabi, Pedram
- Nature Publishing Group UK
Ultracompact 3D microfluidics for time-resolved structural biology
text, January 2020
- Knoska, Juraj; Adriano, Luigi; Awel, Salah
- Deutsches Elektronen-Synchrotron, DESY, Hamburg
Enzyme intermediates captured “on the fly” by mix-and-inject serial crystallography
journal, May 2018
- Olmos, Jose L.; Pandey, Suraj; Martin-Garcia, Jose M.
- BMC Biology, Vol. 16, Issue 1
Megahertz serial crystallography
text, January 2018
- Wiedorn, Max Oliver; Oberthür, Dominik; Bean, Richard
- Nature Publishing Group UK
De novo protein structure determination by heavy-atom soaking in lipidic cubic phase and SIRAS phasing using serial synchrotron crystallography
text, January 2018
- Botha, S.; Baitan, D.; Jungnickel, K. E. J.
- Deutsches Elektronen-Synchrotron, DESY, Hamburg
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