Journal article
Regulation of CFTR Cl− channel gating by ATP binding and hydrolysis
Proceedings of the National Academy of Sciences - PNAS, Vol.97(15), pp.8675-8680
07/18/2000
DOI: 10.1073/pnas.140220597
PMCID: PMC27007
PMID: 10880569
Abstract
Opening and closing of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl
−
channel is regulated by the interaction of ATP with its two cytoplasmic nucleotide-binding domains (NBD). Although ATP hydrolysis by the NBDs is required for normal gating, the influence of ATP binding versus hydrolysis on specific steps in the gating cycle remains uncertain. Earlier work showed that the absence of Mg
2+
prevents hydrolysis. We found that even in the absence of Mg
2+
, ATP could support channel activity, albeit at a reduced level compared with the presence of Mg
2+
. Application of ATP with a divalent cation, including the poorly hydrolyzed CaATP complex, increased the rate of opening. Moreover, in CFTR variants with mutations that disrupt hydrolysis, ATP alone opened the channel and Mg
2+
further enhanced ATP-dependent opening. These data suggest that ATP alone can open the channel and that divalent cations increase ATP binding. Consistent with this conclusion, when we mutated an aspartate thought to bind Mg
2+
, divalent cations failed to increase activity compared with ATP alone. Two observations suggested that divalent cations also stabilize the open state. In wild-type CFTR, CaATP generated a long duration open state, whereas ATP alone did not. With a CFTR variant in which hydrolysis was disrupted, MgATP, but not ATP alone, produced long openings. These results suggest a gating cycle for CFTR in which ATP binding opens the channel and either hydrolysis or dissociation leads to channel closure. In addition, the data suggest that ATP binding and hydrolysis by either NBD can gate the channel.
Details
- Title: Subtitle
- Regulation of CFTR Cl− channel gating by ATP binding and hydrolysis
- Creators
- Mutsuhiro Ikuma - Howard Hughes Medical Institute, Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, IA 52242Michael J Welsh - Howard Hughes Medical Institute, Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, IA 52242
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.97(15), pp.8675-8680
- DOI
- 10.1073/pnas.140220597
- PMID
- 10880569
- PMCID
- PMC27007
- NLM abbreviation
- Proc Natl Acad Sci U S A
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Publisher
- National Academy of Sciences
- Language
- English
- Date published
- 07/18/2000
- Academic Unit
- Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Neurosurgery; Internal Medicine
- Record Identifier
- 9984013999602771
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