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Demonstration af proteolytiske Aktivering af Epithelial Natrium Channel (ENAC) ved at kombinere strømmålinger med påvisning af spaltningsfragmenter
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 af proteolytiske Aktivering af Epithelial Natrium Channel (ENAC) ved at kombinere strømmålinger med påvisning af spaltningsfragmenter

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

Proteolytisk aktivering af epithelial natrium-kanal (ENAC) heterologt udtrykt i Xenopus laevis-oocytter kan påvises ved at kombinere de aktuelle målinger med en biotinylering fremgangsmåde til at undersøge forekomsten af ionkanal spaltningsprodukter på celleoverfladen. Funktionelt vigtige spaltningssteder kan identificeres ved hjælp af steddirigeret mutagenese.

Tags

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

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