What did the Scientists Discover?
Our team has developed a new serial crystallography method for rapid serial data collection with oscillation, lowering the total number of crystals needed. Crystals are deposited onto a silicon chip with thousands of microwells in precise locations. Using a piezoelectric translation stage, each crystal-containing microwell can be positioned in front of the X-ray beam rapidly and precisely. With oscillation, the total amount of data per crystal that can be obtained is improved, without effects from radiation damage. Optimizing data collection by minimizing the effects of background X-ray scatter by air was also paramount.
Broader Impacts of this work?
This work should enable structural biologists to more easily access serial crystallographic techniques, and importantly, simplify room temperature data collection. This in turn should have a dramatic effect on our understanding of protein folding and dynamics under physiological conditions.
Why is this important?
Serial crystallography has much untapped potential within the broader structural biology community, but access to simple and readily available techniques is lacking. Further, economical use of protein crystals- by maximizing the amount of data that can be obtained from each crystal- will make the technique more attractive to users as a whole.
Why did this research need CHESS
MacCHESS has the flexibility needed to design beamlines tailored to user needs. At beamline ID7B2- FlexX, we designed a serial crystallography diffractometer tailor-made for this experiment, and deployed it in a couple of days. This level of rapidity is unique to MacCHESS among other beamlines in the country. We are equally capable of running users with unique needs as well as those who require routine data collection.
Collaborators:
Jennifer L. Wierman, MacCHESS, Cornell University
Olivier Paré-Labrosse, Departments of Chemistry & Physics, University of Toronto and Max Planck Institute for the Structure and Dynamics of Matter, Hamburg, Germany
Antoine Sarracini, Departments of Chemistry & Physics, University of Toronto
Jessica Besaw, Departments of Chemistry & Physics, University of Toronto
Michael J. Cook, MacCHESS, Cornell University
Saeed Oghbaey, Departments of Chemistry & Physics, University of Toronto
Hazem Daoud, Departments of Chemistry & Physics, University of Toronto
Pedram Mehrabi, Max Planck Institute for the Structure and Dynamics of Matter, Hamburg, Germany
Irina Kriksunov, MacCHESS, Cornell University
Anling Kuo, Departments of Biochemistry and Molecular Genetics, University of Toronto
David J. Schuller, MacCHESS, Cornell University
Scott Smith, MacCHESS, Cornell University
Oliver P. Ernst, Departments of Biochemistry and Molecular Genetics, University of Toronto
Doletha M. E. Szebenyi, MacCHESS, Cornell University
Sol M. Gruner, MacCHESS, Department of Physics and Kavli Institute for Nanoscale Science, Cornell University
R. J. Dwayne Miller, Departments of Chemistry & Physics, University of Toronto
Aaron D. Finke, MacCHESS, Cornell University
Publication citation:
Jennifer L. Wierman, Olivier Paré-Labrosse, Antoine Sarracini, Jessica Besaw, Michael J. Cook, Saeed Oghbaey, Hazem Daoud, Pedram Mehrabi, Irina Kriksunov, Anling Kuo, David J. Schuller, Scott Smith, Oliver P. Ernst, Doletha M. E. Szebenyi, Sol M. Gruner, R. J. Dwayne Miller, Aaron D. Finke. Fixed-Target Serial Oscillation Crystallography at Room Temperature. IUCrJ 6 (2019) doi: 10.1107/S2052252519001453
Funding:
This work is based upon research conducted at the Cornell High-Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation and the National Institutes of Health/National Institute of General Medical Sciences under NSF award DMR-1332208, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by award GM-103485 from the National Institutes of Health, through its National Institute of General Medical Sciences. Additional funding was provided by the Max Planck Society for the development of the chip technology, beamline translation system and participation at the beamline. The research was further supported by the Canada Excellence Research Chairs Program (to OPE).
