Well-based crystallization of lipidic cubic phase microcrystals for serial X-ray crystallography experiments

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  • nj5285

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DOI

  • Rebecka Andersson, Göteborgs Universitet
  • ,
  • Cecilia Safari, Göteborgs Universitet
  • ,
  • Petra Bath, Göteborgs Universitet
  • ,
  • Robert Bosman, Göteborgs Universitet
  • ,
  • Anastasya Shilova, Lund University
  • ,
  • Peter Dahl, Göteborgs Universitet
  • ,
  • Swagatha Ghosh, Göteborgs Universitet
  • ,
  • Andreas Dunge, Göteborgs Universitet, AstraZeneca Sweden
  • ,
  • Rasmus Kjeldsen-Jensen
  • ,
  • Jie Nan, Lund University
  • ,
  • Robert L. Shoeman, Max Planck Institute for Medical Research
  • ,
  • Marco Kloos, Max Planck Institute for Medical Research
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  • R. Bruce Doak, Max Planck Institute for Medical Research
  • ,
  • Uwe Mueller, Lund University
  • ,
  • Richard Neutze, Göteborgs Universitet
  • ,
  • Gisela Brändén, Göteborgs Universitet

Serial crystallography is having an increasing impact on structural biology. This emerging technique opens up new possibilities for studying protein structures at room temperature and investigating structural dynamics using time-resolved X-ray diffraction. A limitation of the method is the intrinsic need for large quantities of well ordered micrometre-sized crystals. Here, a method is presented to screen for conditions that produce microcrystals of membrane proteins in the lipidic cubic phase using a well-based crystallization approach. A key advantage over earlier approaches is that the progress of crystal formation can be easily monitored without interrupting the crystallization process. In addition, the protocol can be scaled up to efficiently produce large quantities of crystals for serial crystallography experiments. Using the well-based crystallization methodology, novel conditions for the growth of showers of microcrystals of three different membrane proteins have been developed. Diffraction data are also presented from the first user serial crystallography experiment performed at MAX IV Laboratory.

Original languageEnglish
JournalActa crystallographica Section D: Structural biology
Volume75
Pages (from-to)937-946
Number of pages10
ISSN2059-7983
DOIs
Publication statusPublished - 2019

    Research areas

  • lipidic cubic phase, membrane proteins, protein crystallization, serial crystallography

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