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A Step-by-step Method for the Reconstitution of an ABC Transporter into Nanodisc Lipid Particles
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Biologia
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JoVE Journal Biologia
A Step-by-step Method for the Reconstitution of an ABC Transporter into Nanodisc Lipid Particles

A Step-by-step Method for the Reconstitution of an ABC Transporter into Nanodisc Lipid Particles

DOI:
10.3791/3910-v

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12:25 min

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August 31, 2012

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, ,
1Department of Biochemistry and Molecular Biology,University of British Columbia

Capitoli

  • 00:05Titolo
  • 02:08Preparation of the Membrane Scaffold Protein, MSP
  • 03:53Preparation of the MalFGK2 Complex
  • 05:09Preparation of Phospholipids
  • 06:07Preparation of Bio-Beads & Nanodisc Reconstitution
  • 07:59Nanodisc Analysis: Native Gel Electrophoresis
  • 08:26Nanodisc Analysis: Dynamic Light Scattering (DLS)
  • 09:11Nanodisc Analysis: ATPase Measurements
  • 09:47Results: Activity of the maltose Transporter Reconstituted into Nanodiscs
  • 11:15Conclusion

Summary

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Traduzione automatica

Nanodiscs are small discoid particles that incorporate membrane proteins into a small patch of phospholipid bilayer. We provide a visual protocol that shows the step-by-step incorporation of the MalFGK2 transporter into a disc.

Tags

Nanodisc Lipid ParticlesReconstitution MethodABC TransporterMembrane ProteinsLigand-receptor InteractionsAmphipathic PropertiesApolipoprotein A1Hydrophobic InteractionsDetergent RemovalProtein-to-lipid Ratio

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