Cyclic AMP-dependent protein kinase activation of cystic fibrosis transmembrane conductance regulator chloride channels in planar lipid bilayers

B C Tilly; M C Winter; L S Ostedgaard; C O'Riordan; A E Smith; M J Welsh

doi:10.1016/S0021-9258(19)50113-2

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Cyclic AMP-dependent protein kinase activation of cystic fibrosis transmembrane conductance regulator chloride channels in planar lipid bilayers

Journal article

Open access

Peer reviewed

Cyclic AMP-dependent protein kinase activation of cystic fibrosis transmembrane conductance regulator chloride channels in planar lipid bilayers

B C Tilly, M C Winter, L S Ostedgaard, C O'Riordan, A E Smith and M J Welsh

The Journal of biological chemistry, Vol.267(14), pp.9470-9473

05/15/1992

DOI: 10.1016/S0021-9258(19)50113-2

PMID: 1374403

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Abstract

Membrane vesicles, prepared from mouse NIH-3T3 fibroblasts and Chinese hamster ovary cells expressing high levels of cystic fibrosis transmembrane conductance regulator (CFTR), were fused with Mueller-Rudin planar lipid bilayers. Upon addition of the catalytic subunit of cAMP-dependent protein kinase and ATP, low conductance Cl(-)-selective ion channels were observed in 10 of 16 experiments. The channels had a linear current-voltage relationship and a unitary conductance of approximately 6.5 pS. The channels were more permeable to Cl- than to I- and showed no appreciable time-dependent voltage activation. In contrast, addition of cAMP-dependent protein kinase and ATP to lipid bilayers fused with vesicles prepared from mock transfected (n = 14) cells failed to activate Cl- channels. These data support the conclusion that CFTR is a Cl- channel. They indicate that it can be reconstituted in a planar lipid bilayer and that the biophysical and regulatory properties are very similar to those observed in the native cell membrane. These data also argue against the requirement for loosely associated factors for regulation or function of the channel.

Transfection

Protein Kinases - metabolism

Cricetinae

Membrane Proteins - isolation & purification

Molecular Weight

Membrane Fusion

Membrane Proteins - genetics

Humans

Ion Channels - physiology

Cystic Fibrosis Transmembrane Conductance Regulator

Animals

Chloride Channels

Lipid Bilayers

Membrane Proteins - physiology

Cystic Fibrosis - genetics

Potassium Chloride - pharmacology

Membrane Lipids - metabolism

Membrane Proteins - drug effects

Cell Membrane - metabolism

Membrane Proteins - metabolism

Mice

3T3 Cells

CHO Cells

Details

Title: Subtitle: Cyclic AMP-dependent protein kinase activation of cystic fibrosis transmembrane conductance regulator chloride channels in planar lipid bilayers
Creators: B C Tilly - Howard Hughes Medical Institute, Department of Internal Medicine and Physiology, University of Iowa College of Medicine, Iowa City 52242
M C Winter
L S Ostedgaard
C O'Riordan
A E Smith
M J Welsh
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.267(14), pp.9470-9473
DOI: 10.1016/S0021-9258(19)50113-2
PMID: 1374403
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Language: English
Date published: 05/15/1992
Academic Unit: Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Anatomy and Cell Biology; Fraternal Order of Eagles Diabetes Research Center; Neurosurgery; Internal Medicine
Record Identifier: 9984020716602771

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