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A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl– Channels and Transporters
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A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl– Channels and Transporters

A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl– Channels and Transporters

DOI:
10.3791/52369-v

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07:47 min

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April 20, 2015

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,
1Department of Anesthesiology,Weill Cornell Medical College, 2Department of Physiology and Biophysics,Weill Cornell Medical College, 3Department of Biochemistry,Weill Cornell Medical College

Chapters

  • 00:05Title
  • 01:50Recording Set Up
  • 02:38Preparation of Unilamellar Vesicles
  • 04:10Efflux Measurement
  • 05:27Results: Determination of the Unitary Transport Rate of a CLC-type H+/Cl– Exchanger
  • 07:11Conclusion

Summary

Automatic Translation

Automatic Translation

Proteoliposomes are used to study purified channels and transporters reconstituted in a well-defined biochemical environment. An experimental procedure to measure efflux mediated by these proteins is illustrated. The steps to prepare proteoliposomes, perform the recordings, and analyze data to quantitatively determine the functional properties of the reconstituted protein are described.

Tags

Keywords: ProteoliposomeEfflux AssayCl- ChannelsTransportersMembrane ProteinsIon Sensitive ElectrodeCLC-ec1H+/Cl- ExchangerIon TransportQuantitative AnalysisStructure-function StudiesSmall Molecule EffectsMembrane Composition Effects

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