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Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
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Journal JoVE Biologie
Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

DOI:
10.3791/52427-v

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09:59 min

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March 09, 2015

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, ,
1Programme in Molecular Structure and Function,Hospital for Sick Children, 2Department of Biochemistry,University of Toronto, 3Department of Physiology,University of Toronto

Chapitres

  • 00:05Titre
  • 02:05Reconstitution of CFTR in Liposomes
  • 04:22Measurement of the Iodide Efflux from Reconstituted CFTR in Liposomes
  • 07:29Results: Modulation of CFTR Activity by Small Molecule Inhibitors and Potentiators
  • 08:54Conclusion

Summary

Traduction automatique

Traduction automatique

Described here is a rapid and effective procedure for functional reconstitution of purified wild-type and mutant CFTR protein that preserves activity for this chloride channel, which is defective in Cystic Fibrosis. Iodide efflux from reconstituted proteoliposomes mediated by CFTR allows studies of channel activity and the effects of small molecules.

Tags

CFTRCystic FibrosisChannel ActivityFunctional ReconstitutionProteoliposomesFlux-based AssayF508delG551DPhosphorylationNucleotidesPotentiatorsInhibitors

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