Sign In

JoVE Journal

Bioengineering

Author Produced

A subscription to JoVE is required to view this content.

Construction of Model Lipid Membranes Incorporating G-protein Coupled Receptors (GPCRs)

3.3K Views

09:45 min

February 5th, 2022

February 5th, 2022

3,324 Views

0:05

Introduction

1:16

Preparation of Materials and Reagents

2:58

Hydrogel Preparation

3:40

Agarose and Protein Mixing

4:05

Hydrogel and Lipid Deposition

6:09

Swelling and Harvesting of Vesicles

8:03

Representative Results: Microscopy of Vesicles

8:51

Conclusion

Transcript

Studying integral membrane proteins in their native environment has been a challenge due to the complexity of the plasma membrane.Giant unilamellar lipid vesicles, or GUVs, are one of the most prominent platforms to study biomembrane properties an

Sign in or start your free trial to access this content

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.

We use cookies to enhance your experience on our website.

By continuing to use our website or clicking “Continue”, you are agreeing to accept our cookies.

Learn More