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Detergent-free Ultrafast Reconstitution of Membrane Proteins into Lipid Bilayers Using Fusogenic Complementary-charged Proteoliposomes.
Journal JoVE
Biochimie
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Journal JoVE Biochimie
Detergent-free Ultrafast Reconstitution of Membrane Proteins into Lipid Bilayers Using Fusogenic Complementary-charged Proteoliposomes.

Detergent-free Ultrafast Reconstitution of Membrane Proteins into Lipid Bilayers Using Fusogenic Complementary-charged Proteoliposomes.

DOI:
10.3791/56909-v

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11:10 min

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April 05, 2018

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, , , ,
1First Pavlov State Medical University, 2Clarendon Laboratory, Department of Physics,Oxford University, 3Department of Biological Sciences,Southern Methodist University

Chapitres

  • 00:04Titre
  • 01:16Formation of Fusogenic GUV by Inverted Emulsion Method
  • 05:59Delivery of Membrane Proteins and Assembly of Electron Transport Chain in the LUV and GUV followed by ATP Production
  • 08:21Results: Use of Vesicle Fusion and ACMA Quenching Assay to Analyze the Assembly of Complex Multi-component Membrane Systems
  • 10:11Conclusion

Summary

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Here we present two ultrafast protocols for reconstitution of membrane proteins into fusogenic proteoliposomes, and fusion of such proteoliposomes with target lipid bilayers for detergent-free delivery of these membrane proteins into the postfusion bilayer. The combination of these approaches enables fast and easily controlled assembly of complex multi-component membrane systems.

Tags

Keywords: Membrane ProteinsLipid BilayersFusogenic ProteoliposomesDetergent-freeReconstitutionArtificial CellMembrane TransportSynthetic Biology

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