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