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Title: 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:
ORCiD logo [1];  [1]; ORCiD logo [1];  [1];  [1]; ORCiD logo [2];  [3]; ORCiD logo [3]; ORCiD logo [4]; ORCiD logo [5];  [6]; ORCiD logo [3]; ORCiD logo [1];  [1]; ORCiD logo [1];  [1];  [1]; ORCiD logo [1]; ORCiD logo [3]; ORCiD logo [7] more »; ORCiD logo [8];  [1]; ORCiD logo [1];  [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [3];  [1]; ORCiD logo [1];  [1];  [9];  [1]; ORCiD logo [3]; ORCiD logo [1]; ORCiD logo [1] « less
  1. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  2. California State Univ., Northridge, CA (United States)
  3. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  4. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Norwegian Univ. of Science and Technology, Trondheim (Norway)
  5. Uppsala Univ., Uppsala (Sweden)
  6. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); European X-ray Free-Electron Laser Facility GmbH (XFEL), Schenefeld (Germany)
  7. Univ. of Oxford, Oxford (England)
  8. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  9. 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. 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, and Oberthuer, Dominik. Mon . "Mix-and-diffuse serial synchrotron crystallography". United States. doi: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}
}

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Cited by: 13 works
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Figures / Tables:

Figure 1 Figure 1: Schematics of the experimental setup. (a) The native protein crystal suspension (yellow) is mixed inline with the substrate solution (blue) before being deposited onto the detector side of the tape. The X-ray beam (red) is focused on the centre of the polyimide tape (yellow ribbon), which is beingmore » drawn from the feeder roll on the left to the collector roll on the right. A diffraction pattern measured in transmission is illustrated as red dots on the grey detector. (b) A technical drawing of the tape-drive device shows how tape is held under tension between two rollers near the interaction region and that X-rays pass through a hole in the centre of the device.« less

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Mix-and-diffuse serial synchrotron crystallography
text, January 2017

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  • DOI: 10.3204/pubdb-2017-10745

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    • DOI: 10.3204/pubdb-2018-03102

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    • DOI: 10.3204/pubdb-2019-01297

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