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Protein-Eye View of the in Meso Crystallization Mechanism.
Journal article

Protein-Eye View of the in Meso Crystallization Mechanism.

  • van T Hag L Department of Health Sciences and Technology , ETH Zurich , CH-8092 Zurich , Switzerland.
  • de Campo L School of Science, College of Science, Engineering and Health , RMIT University , Melbourne , Victoria 3001 , Australia.
  • Tran N Structural Biology Division , The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria 3052 , Australia.
  • Sokolova A Structural Biology Division , The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria 3052 , Australia.
  • Trenker R Structural Biology Division , The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria 3052 , Australia.
  • Call ME Scientific Activities Division , European Spallation Source ERIC , Lund 224 84 , Sweden.
  • Call MJ School of Science, College of Science, Engineering and Health , RMIT University , Melbourne , Victoria 3001 , Australia.
  • Garvey CJ School of Science, College of Science, Engineering and Health , RMIT University , Melbourne , Victoria 3001 , Australia.
  • Leung AE Department of Health Sciences and Technology , ETH Zurich , CH-8092 Zurich , Switzerland.
  • Darwish TA School of Science, College of Science, Engineering and Health , RMIT University , Melbourne , Victoria 3001 , Australia.
  • Krause-Heuer A
  • Knott R
  • Meikle TG
  • Drummond CJ
  • Mezzenga R
  • Conn CE
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  • 2019-05-25
Published in:
  • Langmuir : the ACS journal of surfaces and colloids. - 2019
Crystallization
Glycerides
Lipid Bilayers
X-Ray Diffraction
English For evolving biological and biomedical applications of hybrid protein?lipid materials, understanding the behavior of the protein within the lipid mesophase is crucial. After more than two decades since the invention of the in meso crystallization method, a protein-eye view of its mechanism is still lacking. Numerous structural studies have suggested that integral membrane proteins preferentially partition at localized flat points on the bilayer surface of the cubic phase with crystal growth occurring from a local fluid lamellar L? phase conduit. However, studies to date have, by necessity, focused on structural transitions occurring in the lipid mesophase. Here, we demonstrate using small-angle neutron scattering that the lipid bilayer of monoolein (the most commonly used lipid for in meso crystallization) can be contrast-matched using deuteration, allowing us to isolate scattering from encapsulated peptides during the crystal growth process for the first time. During in meso crystallization, a clear decrease in form factor scattering intensity of the peptides was observed and directly correlated with crystal growth. A transient fluid lamellar L? phase was observed, providing direct evidence for the proposed mechanism for this technique. This suggests that the peptide passes through a transition from the cubic QII phase, via an L? phase to the lamellar crystalline Lc phase with similar layered spacing. When high protein loading was possible, the lamellar crystalline Lc phase of the peptide in the single crystals was observed. These findings show the mechanism of in meso crystallization for the first time from the perspective of integral membrane proteins.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/160524
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