2
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals

      research-article
      a , b , b , c , d , c , e , f , g , h , g , i , b , j , d , c , a , b , d , k , l , k , m , n , h , d , g , o , p , g , g , q , r , g , h , g , h , b , s , t , b , s , u , v , g , q , d , g , h , e , f , w , c , b , b , *
      IUCrJ
      International Union of Crystallography
      microcrystals, batch crystallization, serial crystallography, MicroED

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Serial X-ray crystallography and microcrystal electron diffraction were performed using microcrystals of the enzyme cyclophilin A. The results highlight the strengths and weakness of the two complementary methods.

          Abstract

          Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometre to micrometre scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges that are not encountered in traditional macromolecular crystallography experiments. Here, XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A are described. The results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample-preparation and delivery methods required for each type of experiment affect the crystal structure of the enzyme.

          Related collections

          Author and article information

          Journal
          IUCrJ
          IUCrJ
          IUCrJ
          IUCrJ
          International Union of Crystallography
          2052-2525
          01 March 2020
          26 February 2020
          26 February 2020
          : 7
          : Pt 2 ( publisher-idID: m200200 )
          : 306-323
          Affiliations
          [a ]Graduate Program in Biophysics, University of California, San Francisco , San Francisco, California, USA
          [b ]Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco , San Francisco, California, USA
          [c ]Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory , Berkeley, California, USA
          [d ]Linac Coherent Light Source, SLAC National Accelerator Laboratory , Menlo Park, California, USA
          [e ]Howard Hughes Medical Institute, University of California, Los Angeles , Los Angeles, California, USA
          [f ]Department of Biological Chemistry, University of California, Los Angeles , Los Angeles, California, USA
          [g ] RIKEN SPring-8 Center , 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan
          [h ]Department of Cell Biology, Graduate School of Medicine, Kyoto University , Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan
          [i ]Department of Biological Science, Graduate School of Science, The University of Tokyo , Tokyo, Japan
          [j ]Laboratory for Drug Discovery, Pharmaceuticals Research Center, Asahi Kasei Pharma Corporation , 632-1 Mifuku, Izunokuni-shi, Shizuoka 410-2321, Japan
          [k ]SSRL, SLAC National Accelerator Laboratory , Menlo Park, California, USA
          [l ]Department of Biology, San Francisco State University , San Francisco, California, USA
          [m ]Department of Chemistry and Biotechnology, Graduate School of Engineering, Tottori University , 4-101 Koyama-cho, Minami, Tottori 680-8552, Japan
          [n ]Center for Research on Green Sustainable Chemistry, Tottori University , Tottori, Japan
          [o ]Graduate School of Life Science, University of Hyogo , Ako-gun, Hyogo 678-1297, Japan
          [p ]Department of Chemistry, New York University , New York, USA
          [q ] Japan Synchrotron Radiation Research Institute , 1-1-1 Kouto, Sayo, Hyogo 679-5198, Japan
          [r ]Institute for Neurodegenerative Diseases, University of California, San Francisco , San Francisco, California, USA
          [s ]Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco , San Francisco, California, USA
          [t ]Bioscience Department, SLAC National Accelerator Laboratory , Menlo Park, California, USA
          [u ]Structural Biology Research Center, Institute of Materials Structure Science, KEK/High Energy Accelerator Research Organization , Tsukuba, Ibaraki 305-0034, Japan
          [v ]Department of Biology and Biological Engineering, California Institute of Technology , Pasadena, California, USA
          [w ]Department of Physiology, University of California, Los Angeles , Los Angeles, California, USA
          Author notes
          Correspondence e-mail: mct.ucsf@ 123456gmail.com
          [‡]

          Present address: MRC Laboratory of Molecular Biology, Cambridge, England.

          Author information
          https://orcid.org/0000-0003-0474-7673
          https://orcid.org/0000-0001-9851-7355
          https://orcid.org/0000-0003-2697-2767
          https://orcid.org/0000-0002-5292-4470
          https://orcid.org/0000-0003-2414-9427
          https://orcid.org/0000-0002-6820-0936
          https://orcid.org/0000-0002-1451-7612
          https://orcid.org/0000-0003-1062-4792
          https://orcid.org/0000-0003-2358-841X
          https://orcid.org/0000-0002-9592-397X
          https://orcid.org/0000-0003-1218-3759
          https://orcid.org/0000-0003-3928-1244
          https://orcid.org/0000-0002-5080-2859
          https://orcid.org/0000-0002-6099-2027
          Article
          mf5038 IUCRAJ S205225252000072X
          10.1107/S205225252000072X
          7055375
          32148858
          67ec72da-81fc-4c20-a893-40b19fadea1e
          © Alexander M. Wolff et al. 2020

          This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.

          History
          : 13 September 2019
          : 21 January 2020
          Page count
          Pages: 18
          Funding
          Funded by: National Science Foundation, BioXFEL Science and Technology Center
          Award ID: STC-1231306
          Award Recipient : Michael Thompson Award Recipient : James Fraser
          Funded by: National Institutes of Health, National Heart, Lung, and Blood Institute
          Award ID: F32 HL129989
          Award Recipient : Michael Thompson
          Funded by: National Institutes of Health, National Institute of General Medical Sciences
          Award ID: NIH GM123159
          Award ID: NIH GM124149
          Award ID: NIH GM117126
          Award Recipient : James Fraser Award Recipient : James Fraser Award Recipient : Nicholas Sauter
          Funded by: Office of the President, University of California
          Award ID: LFR-17-476732
          Award Recipient : James Fraser
          Funded by: U.S. Department of Energy, Office of Science
          Award ID: DE-AC02-76SF00515
          Award ID: DEAC02-05CH11231
          Funded by: David and Lucile Packard Foundation
          Funded by: Japan Agency for Medical Research and Development
          Funded by: University of California, San Francisco
          This work was funded by National Science Foundation, BioXFEL Science and Technology Center grant STC-1231306 to Michael Thompson and James Fraser. National Institutes of Health, National Heart, Lung, and Blood Institute grant F32 HL129989 to Michael Thompson. National Institutes of Health, National Institute of General Medical Sciences grants NIH GM123159, NIH GM124149, and NIH GM117126. Office of the President, University of California grant LFR-17-476732 to James Fraser. U.S. Department of Energy, Office of Science grants DE-AC02-76SF00515 and DEAC02-05CH11231. David and Lucile Packard Foundation grant . Japan Agency for Medical Research and Development grant . University of California, San Francisco grant .
          Categories
          Research Papers

          microcrystals,batch crystallization,serial crystallography,microed

          Comments

          Comment on this article