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Construction of Model Lipid Membranes Incorporating G-protein Coupled Receptors (GPCRs)
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Construction of Model Lipid Membranes Incorporating G-protein Coupled Receptors (GPCRs)
DOI:

09:45 min

February 05, 2022

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Chapters

  • 00:05Introduction
  • 01:16Preparation of Materials and Reagents
  • 02:58Hydrogel Preparation
  • 03:40Agarose and Protein Mixing
  • 04:05Hydrogel and Lipid Deposition
  • 06:09Swelling and Harvesting of Vesicles
  • 08:03Representative Results: Microscopy of Vesicles
  • 08:51Conclusion

Summary

Automatic Translation

This protocol utilizes agarose swelling as a powerful and generalizable technique for incorporating integral membrane proteins (IMPs) into giant unilamellar lipid vesicles (GUVs), as described here for the reconstitution of the human 1A serotonin receptor protein (5-HT1AR), one of the classes of pharmacologically important G protein-coupled receptors.

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