Two-Dimensional Crystallization of Membrane Proteins by Reconstitution Through Dialysis

  • Matthew C. Johnson
  • Tina M. Dreaden
  • Laura Y. Kim
  • Frederik Rudolph
  • Bridgette A. Barry
  • Ingeborg Schmidt-Krey
Part of the Methods in Molecular Biology book series (MIMB, volume 955)


Studies of membrane proteins by two-dimensional (2D) crystallization and electron crystallography have provided crucial information on the structure and function of a rapidly growing number of these intricate proteins within a close-to-native lipid bilayer. Here we provide protocols for planning and executing 2D crystallization trials by detergent removal through dialysis, including the preparation of phospholipids and the dialysis setup. General factors to be considered, such as the protein preparation, solubilizing detergent, lipid for reconstitution, and buffer conditions are discussed. Several 2D crystallization conditions are highlighted that have shown great promise to grow 2D crystals within a surprisingly short amount of time. Finally, conditions for optimizing order and size of 2D crystals are outlined.

Key words

Two-dimensional crystallization 2D crystals Membrane protein Dialysis Lipid Detergent Electron crystallography 



We wish to thank our colleagues in the laboratory as well as in the cryo-EM community for helpful discussions over the years. We are particularly indebted to our collaborators for entrusting us with their valuable samples. Research in the Schmidt-Krey laboratory contributing to some of these observations was funded by NIH grant HL090630 and the Integrative Biosystems Institute.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Matthew C. Johnson
    • 1
  • Tina M. Dreaden
    • 2
  • Laura Y. Kim
    • 1
  • Frederik Rudolph
    • 1
  • Bridgette A. Barry
    • 2
  • Ingeborg Schmidt-Krey
    • 3
  1. 1.School of BiologyGeorgia Institute of TechnologyAtlantaUSA
  2. 2.School of Chemistry and BiochemistryGeorgia Institute of TechnologyAtlantaUSA
  3. 3.School of Biology, School of Chemistry and BiochemistryGeorgia Institute of TechnologyAtlantaUSA

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