Abstract
The amiloride-sensitive epithelial Na+ channel (ENaC) is typically composed of three structurally related subunits termed alpha, beta, and gamma. We describe methods to determine the functional subunit stoichiometry of ENaC based on a biophysical approach that was first introduced in 1991 to determine the subunit stoichiometry of a voltage-gated K+ channel. The strategy is to analyze channel sensitivity to a specific blocker when various mixtures of block-sensitive and blocker-insensitive subunits are coexpressed in a heterologous expression system. Details related to the expression of wild type and mutant ENaCs in Xenopus oocytes and the examination of blocker sensitivity by two-electrode voltage clamp, as well as analysis of data are provided.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 53-63 |
| Number of pages | 11 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 337 |
| State | Published - 2006 |
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ASJC Scopus subject areas
- Medicine(all)
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Biophysical approach to determine the subunit stoichiometry of the epithelial sodium channel using the Xenopus laevis oocyte expression system. / Kosari, Farhad; Sheng, Shaohu; Kleyman, Thomas R.
In: Methods in molecular biology (Clifton, N.J.), Vol. 337, 2006, p. 53-63.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Biophysical approach to determine the subunit stoichiometry of the epithelial sodium channel using the Xenopus laevis oocyte expression system.
AU - Kosari, Farhad
AU - Sheng, Shaohu
AU - Kleyman, Thomas R.
PY - 2006
Y1 - 2006
N2 - The amiloride-sensitive epithelial Na+ channel (ENaC) is typically composed of three structurally related subunits termed alpha, beta, and gamma. We describe methods to determine the functional subunit stoichiometry of ENaC based on a biophysical approach that was first introduced in 1991 to determine the subunit stoichiometry of a voltage-gated K+ channel. The strategy is to analyze channel sensitivity to a specific blocker when various mixtures of block-sensitive and blocker-insensitive subunits are coexpressed in a heterologous expression system. Details related to the expression of wild type and mutant ENaCs in Xenopus oocytes and the examination of blocker sensitivity by two-electrode voltage clamp, as well as analysis of data are provided.
AB - The amiloride-sensitive epithelial Na+ channel (ENaC) is typically composed of three structurally related subunits termed alpha, beta, and gamma. We describe methods to determine the functional subunit stoichiometry of ENaC based on a biophysical approach that was first introduced in 1991 to determine the subunit stoichiometry of a voltage-gated K+ channel. The strategy is to analyze channel sensitivity to a specific blocker when various mixtures of block-sensitive and blocker-insensitive subunits are coexpressed in a heterologous expression system. Details related to the expression of wild type and mutant ENaCs in Xenopus oocytes and the examination of blocker sensitivity by two-electrode voltage clamp, as well as analysis of data are provided.
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UR - http://www.scopus.com/inward/citedby.url?scp=39049185619&partnerID=8YFLogxK
M3 - Article
C2 - 16929938
AN - SCOPUS:39049185619
VL - 337
SP - 53
EP - 63
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -