Abstract
Hypertension is a multifactorial disorder that results in an increased risk of cardiovascular and end-stage renal disease. Liddle's disease represents a specific hypertensive disease and expresses itself in the human population as an autosomal dominant trait. Recent experimental evidence indicates that patients with Liddle's disease have constitutively active amiloride-sensitive Na+ channels and that these channels are phenotypically expressed in lymphocytes obtained from normal and affected members of the original Liddle's kindred. Linkage analysis indicates that this disease results from a deletion of the carboxy-terminal region of the β-subunit of a recently cloned epithelial Na+ channel (ENaC). We report the successful immunopurification and reconstitution of both normal and constitutively active lymphocyte Na+ channels into planar lipid bilayers. These channels display all of the characteristics typical of renal Na+ channels, including sensitivity to protein kinase A phosphorylation. We demonstrate that gating of normal Na+ channels is removed by cytoplasmic trypsin digestion and that the constitutively active Liddle's Na+ channels are blocked by a β- or γ- ENaC carboxy-terminal peptide in a GTP-dependent fashion.
Original language | English |
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Pages (from-to) | C214-C223 |
Journal | American Journal of Physiology - Cell Physiology |
Volume | 270 |
Issue number | 1 39-1 |
DOIs | |
Publication status | Published - Jan 1996 |
Keywords
- amiloride
- hypertension
- immunopurification
- planar lipid bilayers
- protein purification
ASJC Scopus subject areas
- Physiology
- Cell Biology