Subunit stoichiometry of a core conduction element in a cloned epithelial amiloride-sensitive Na+ channel

Bakhrom K. Berdiev, Katherine H. Karlson, Biljana Jovov, Pierre Jean Ripoll, Ryan Morris, Dominique Loffing-Cueni, Patricia Halpin, Bruce A. Stanton, Thomas R. Kleyman, Iskander I. Ismailov

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Abstract

The molecular composition of a core conduction element formed by the α- subunit of cloned epithelial Na+ channels (ENaC) was studied in Planar lipid bilayers. Two pairs of in vitro translated proteins were employed in combinatorial experiments: 1) wild-type (WT) and an N-terminally truncated α(ΔN)-rENaC that displays accelerated kinetics (τ(o) = 32 ± 13 ms, τ(o) = 42 ± 11 ms), as compared with the WT channel (τ(c1) = 18 ± 8 ms, τ(c2) = 252 ± 31 ms, and τ(o) = 157 ± 43 ms); and 2) WT and an amiloride binding mutant, α(Δ278-283)-rENaC. The channels that formed in a α(WT):α(AN) mixture fell into two groups: one with τ(o) and τ(c) that corresponded to those exhibited by the α(ΔN)-rENaC alone, and another with a double- exponentially distributed closed time and a single-exponentially distributed open time that corresponded to the α(WT)-rENaC alone. Five channel subtypes with distinct sensitivities to amiloride were found in a 1α(WT):1α(Δ278- 283) protein mixture. Statistical analyses of the distributions of channel phenotypes observed for either set of the WT:mutant combinations suggest a tetrameric organization of α-subunits as a minimal model for the core conduction element in ENaCs.

Original languageEnglish
Pages (from-to)2292-2301
Number of pages10
JournalBiophysical Journal
Volume75
Issue number5
DOIs
Publication statusPublished - Nov 1998

ASJC Scopus subject areas

  • Biophysics

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    Berdiev, B. K., Karlson, K. H., Jovov, B., Ripoll, P. J., Morris, R., Loffing-Cueni, D., Halpin, P., Stanton, B. A., Kleyman, T. R., & Ismailov, I. I. (1998). Subunit stoichiometry of a core conduction element in a cloned epithelial amiloride-sensitive Na+ channel. Biophysical Journal, 75(5), 2292-2301. https://doi.org/10.1016/S0006-3495(98)77673-8