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Demonstration of Proteolytic Activation of the Epithelial Sodium Channel (ENaC) by Combining Current Measurements with Detection of Cleavage Fragments
JoVE Journal
Biology
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JoVE Journal Biology
Demonstration of Proteolytic Activation of the Epithelial Sodium Channel (ENaC) by Combining Current Measurements with Detection of Cleavage Fragments

Demonstration of Proteolytic Activation of the Epithelial Sodium Channel (ENaC) by Combining Current Measurements with Detection of Cleavage Fragments

DOI:
10.3791/51582-v

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08:56 min

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July 05, 2014

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, ,
1Institut für Zelluläre und Molekulare Physiologie,Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU)

Chapters

  • 00:05Title
  • 01:11Two-electrode Voltage-clamp Experiments
  • 02:41Measurement of Amiloride-sensitive Whole-cell Currents
  • 04:17Biotinylation Assay and Detection of ENaC Cleavage Fragments at Cell Surface by Western Blot
  • 07:08Results: Proteolytic Activation of the Epithelial Sodium Channel
  • 08:09Conclusion

Summary

Automatic Translation

Automatic Translation

Proteolytic activation of the epithelial sodium channel (ENaC) heterologously expressed in Xenopus laevis oocytes can be demonstrated by combining current measurements with a biotinylation approach to investigate the appearance of ion channel cleavage products at the cell surface. Functionally important cleavage sites can be identified by using site-directed mutagenesis.

Tags

Proteolytic ActivationEpithelial Sodium Channel (ENaC)Two-electrode Voltage-clampAmiloride-sensitive CurrentProteolytic CleavageBiotinylationGamma-ENaC Fragment

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