Journal
/
Biology
/
Reconstitution of a Kv Channel into Lipid Membranes for Structural and Functional Studies
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Reconstitution of a Kv Channel into Lipid Membranes for Structural and Functional Studies

Reconstitution of a Kv Channel into Lipid Membranes for Structural and Functional Studies

DOI:
10.3791/50436-v

•

10:22 min

•

July 13, 2013

•

, , ,
1Department of Cell Biology,University of Texas Southwestern Medical Center at Dallas

Chapters

  • 00:05Title
  • 01:27Liposome Preparation and Detergent-induced Fusion of Vesicles
  • 04:15Removal of the Detergents to Form Proteoliposomes
  • 06:28Functional Study of the Ion Channel Activities in Black Lipid Membranes
  • 08:49Results: Two-Dimensional Crystallization of KvAP in Membranes
  • 09:56Conclusion

Summary

Automatic Translation

Automatic Translation

Procedures for complete reconstitution of a prototype voltage-gated potassium channel into lipid membranes are described. The reconstituted channels are suitable for biochemical assays, electrical recordings, ligand screening and electron crystallographic studies. These methods may have general applications to the structural and functional studies of other membrane proteins.

Tags

Keywords: Kv ChannelLipid ReconstitutionMembrane ProteinsVoltage-gated Potassium ChannelLipid-protein InteractionMembrane SystemsDetergent-induced Fusion2D Crystals

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/50436?article_type=v

Crystallizing Membrane Proteins for Structure Determination using Lipidic Mesophases
check_url/50436?article_type=v

Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography
check_url/50436?article_type=v

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
check_url/50436?article_type=v

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
check_url/50436?article_type=v

A Step-by-step Method for the Reconstitution of an ABC Transporter into Nanodisc Lipid Particles
check_url/50436?article_type=v

Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases
check_url/50436?article_type=v

Harvesting and Cryo-cooling Crystals of Membrane Proteins Grown in Lipidic Mesophases for Structure Determination by Macromolecular Crystallography
check_url/50436?article_type=v

Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes
check_url/50436?article_type=v

Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies
check_url/50436?article_type=v

Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

Read Article