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De novo protein structure determination by heavy-atom soaking in lipidic cubic phase and SIRAS phasing using serial synchrotron crystallography.
Botha, S; Baitan, D; Jungnickel, K E J; Oberthür, D; Schmidt, C; Stern, S; Wiedorn, M O; Perbandt, M; Chapman, H N; Betzel, C.
Affiliation
  • Botha S; Institute of Biochemistry and Molecular Biology, Chemistry Department, University of Hamburg, Martin-Luther-King Platz 6, 20146 Hamburg, Germany.
  • Baitan D; Laboratory for Structural Biology of Infection and Inflammation, c/o DESY, Building 22a, Notkestrasse 85, 22607 Hamburg, Germany.
  • Jungnickel KEJ; The Hamburg Centre for Ultrafast Imaging, Luruper Chaussee 149, 22761 Hamburg, Germany.
  • Oberthür D; Institute of Biochemistry and Molecular Biology, Chemistry Department, University of Hamburg, Martin-Luther-King Platz 6, 20146 Hamburg, Germany.
  • Schmidt C; Laboratory for Structural Biology of Infection and Inflammation, c/o DESY, Building 22a, Notkestrasse 85, 22607 Hamburg, Germany.
  • Stern S; Xtal Concepts GmbH, Marlowring 19, 22525 Hamburg, Germany.
  • Wiedorn MO; Institute of Biochemistry and Molecular Biology, Chemistry Department, University of Hamburg, Martin-Luther-King Platz 6, 20146 Hamburg, Germany.
  • Perbandt M; Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany.
  • Chapman HN; Institute of Biochemistry and Molecular Biology, Chemistry Department, University of Hamburg, Martin-Luther-King Platz 6, 20146 Hamburg, Germany.
  • Betzel C; Laboratory for Structural Biology of Infection and Inflammation, c/o DESY, Building 22a, Notkestrasse 85, 22607 Hamburg, Germany.
IUCrJ ; 5(Pt 5): 524-530, 2018 Sep 01.
Article in En | MEDLINE | ID: mdl-30224955
ABSTRACT
During the past few years, serial crystallography methods have undergone continuous development and serial data collection has become well established at high-intensity synchrotron-radiation beamlines and XFEL radiation sources. However, the application of experimental phasing to serial crystallography data has remained a challenging task owing to the inherent inaccuracy of the diffraction data. Here, a particularly gentle method for incorporating heavy atoms into micrometre-sized crystals utilizing lipidic cubic phase (LCP) as a carrier medium is reported. Soaking in LCP prior to data collection offers a new, efficient and gentle approach for preparing heavy-atom-derivative crystals directly before diffraction data collection using serial crystallography methods. This approach supports effective phasing by utilizing a reasonably low number of diffraction patterns. Using synchrotron radiation and exploiting the anomalous scattering signal of mercury for single isomorphous replacement with anomalous scattering (SIRAS) phasing resulted in high-quality electron-density maps that were sufficient for building a complete structural model of proteinase K at 1.9 Šresolution using automatic model-building tools.
Key words

Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: IUCrJ Year: 2018 Document type: Article Affiliation country: Germany

Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: IUCrJ Year: 2018 Document type: Article Affiliation country: Germany