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 journalArticlepeer-review

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 1 2004

Keywords

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

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health
  • Pulmonary and Respiratory Medicine

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