Design of ultra-swollen lipidic mesophases for the crystallization of membrane proteins with large extracellular domains

Nat Commun. 2018 Feb 7;9(1):544. doi: 10.1038/s41467-018-02996-5.


In meso crystallization of membrane proteins from lipidic mesophases is central to protein structural biology but limited to membrane proteins with small extracellular domains (ECDs), comparable to the water channels (3-5 nm) of the mesophase. Here we present a strategy expanding the scope of in meso crystallization to membrane proteins with very large ECDs. We combine monoacylglycerols and phospholipids to design thermodynamically stable ultra-swollen bicontinuous cubic phases of double-gyroid (Ia3d), double-diamond (Pn3m), and double-primitive (Im3m) space groups, with water channels five times larger than traditional lipidic mesophases, and showing re-entrant behavior upon increasing hydration, of sequences Ia3d→Pn3m→Ia3d and Pn3m→Im3m→Pn3m, unknown in lipid self-assembly. We use these mesophases to crystallize membrane proteins with ECDs inaccessible to conventional in meso crystallization, demonstrating the methodology on the Gloeobacter ligand-gated ion channel (GLIC) protein, and show substantial modulation of packing, molecular contacts and activation state of the ensued proteins crystals, illuminating a general strategy in protein structural biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Membrane*
  • Crystallization / methods
  • Fatty Acids, Monounsaturated / chemistry
  • Ion Channels
  • Membrane Proteins / chemistry*
  • Phase Transition
  • Phosphatidylglycerols / chemistry*
  • Protein Domains
  • Thermodynamics
  • Water
  • X-Ray Diffraction


  • Fatty Acids, Monounsaturated
  • Ion Channels
  • Membrane Proteins
  • Phosphatidylglycerols
  • Water
  • distearoyl phosphatidylglycerol