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

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)79-83
Number of pages5
JournalJournal of Cystic Fibrosis
Volume3
Issue numberSUPPL. 2
DOIs
Publication statusPublished - Aug 2004
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Chloride Channels
Cyclic Nucleotides
Endoplasmic Reticulum
Cyclic AMP
In Vitro Techniques
Cell Membrane
Technology
Proteins

Keywords

  • Bilayers
  • Channel activity
  • Disease-causing mutants
  • Purification
  • Reconstitution
  • Vesicles

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Benos, D. J., Berdiev, B. K., Ismailov, I. I., Ostedgaard, L. S., Kogan, I., Li, C., ... Bear, C. E. (2004). Methods to study CFTR protein in vitro. Journal of Cystic Fibrosis, 3(SUPPL. 2), 79-83. https://doi.org/10.1016/j.jcf.2004.05.018

Methods to study CFTR protein in vitro. / Benos, Dale J.; Berdiev, Bakhrom K.; Ismailov, Iskander I.; Ostedgaard, Lynda S.; Kogan, Ilana; Li, Canhui; Ramjeesingh, Mohabir; Bear, Christine E.

In: Journal of Cystic Fibrosis, Vol. 3, No. SUPPL. 2, 08.2004, p. 79-83.

Research output: Contribution to journalArticle

Benos, DJ, Berdiev, BK, Ismailov, II, Ostedgaard, LS, Kogan, I, Li, C, Ramjeesingh, M & Bear, CE 2004, 'Methods to study CFTR protein in vitro', Journal of Cystic Fibrosis, vol. 3, no. SUPPL. 2, pp. 79-83. https://doi.org/10.1016/j.jcf.2004.05.018
Benos DJ, Berdiev BK, Ismailov II, Ostedgaard LS, Kogan I, Li C et al. Methods to study CFTR protein in vitro. Journal of Cystic Fibrosis. 2004 Aug;3(SUPPL. 2):79-83. https://doi.org/10.1016/j.jcf.2004.05.018
Benos, Dale J. ; Berdiev, Bakhrom K. ; Ismailov, Iskander I. ; Ostedgaard, Lynda S. ; Kogan, Ilana ; Li, Canhui ; Ramjeesingh, Mohabir ; Bear, Christine E. / Methods to study CFTR protein in vitro. In: Journal of Cystic Fibrosis. 2004 ; Vol. 3, No. SUPPL. 2. pp. 79-83.
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