Methods to study CFTR protein in vitro

Dale J Benos; Bakhrom K Berdiev; Iskander I Ismailov; Lynda S Ostedgaard; Ilana Kogan; Canhui Li; Mohabir Ramjeesingh; Christine E Bear

doi:10.1016/j.jcf.2004.05.018

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Journal article

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Methods to study CFTR protein in vitro

Dale J Benos, Bakhrom K Berdiev, Iskander I Ismailov, Lynda S Ostedgaard, Ilana Kogan, Canhui Li, Mohabir Ramjeesingh and Christine E Bear

Journal of cystic fibrosis, Vol.3(2), pp.79-83

2004

DOI: 10.1016/j.jcf.2004.05.018

PMID: 15463933

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Abstract

CFTR is a cyclic AMP and nucleotide-related chloride-selective channel with a low unitary conductance. Many of the physiological roles of CFTR are effectively studied in intact cells and tissues. However, there are also several clear advantages to the application of cell-free technologies to the study of the biochemical and biophysical properties of CFTR. When expressed in heterologous cells, CFTR is processed relatively poorly, depending, however, on the cell-type analysed. In some cells, only 20–25% of the protein which is initially synthesized exits the endoplasmic reticulum to insert into the cell membrane [Cell 83 (1995) 121; EMBO J. 13 (1994) 6076]. Further, many of the disease-causing mutants of CFTR result in even lower processing efficiencies. Therefore, several procedures have been developed to study regulated CFTR channel function expressed in microsomal membanes and following its purification and reconstitution. These experimental approaches and their application are discussed here.

Vesicles

Reconstitution

Purification

Disease-causing mutants

Channel activity

Bilayers

Details

Title: Subtitle: Methods to study CFTR protein in vitro
Creators: Dale J Benos - Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
Bakhrom K Berdiev - Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
Iskander I Ismailov - Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
Lynda S Ostedgaard - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA
Ilana Kogan - Department of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children and Department of Physiology, University of Toronto, Toronto, Ontario, Canada
Canhui Li - Department of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children and Department of Physiology, University of Toronto, Toronto, Ontario, Canada
Mohabir Ramjeesingh - Department of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children and Department of Physiology, University of Toronto, Toronto, Ontario, Canada
Christine E Bear - Department of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children and Department of Physiology, University of Toronto, Toronto, Ontario, Canada
Resource Type: Journal article
Publication Details: Journal of cystic fibrosis, Vol.3(2), pp.79-83
Publisher: Elsevier B.V
DOI: 10.1016/j.jcf.2004.05.018
PMID: 15463933
ISSN: 1569-1993
eISSN: 1873-5010
Language: English
Date published: 2004
Academic Unit: Internal Medicine
Record Identifier: 9984094546102771

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