Altered chloride ion channel kinetics associated with the delta F508 cystic fibrosis mutation

Nature. 1991 Dec 19-26;354(6354):526-8. doi: 10.1038/354526a0.

Abstract

Cystic fibrosis is associated with a defect in epithelial chloride ion transport which is caused by mutations in a membrane protein called CFTR (cystic fibrosis transmembrane conductance regulator). Heterologous expression of CFTR produces cyclicAMP-sensitive Cl(-)-channel activity. Deletion of phenylalanine at amino-acid position 508 in CFTR (delta F508 CFTR) is the most common mutation in cystic fibrosis. It has been proposed that this mutation prevents glycoprotein maturation and its transport to its normal cellular location. We have expressed both CFTR and delta F508 CFTR in Vero cells using recombinant vaccinia virus. Although far less delta F508 CFTR reached the plasma membrane than normal CFTR, sufficient delta F508 CFTR was expressed at the plasma membrane to permit functional analysis. delta F508 CFTR expression induced a reduced activity of the cAMP-activated Cl- channel, with conductance, anion selectivity and open-time kinetics similar to those of CFTR, but with much greater closed times, resulting in a large decrease of open probability. The delta F508 mutation thus seems to have two major consequences, an abnormal translocation of the CFTR protein which limits membrane insertion, and an abnormal function in mediating Cl- transport.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Chloride Channels
  • Cyclic AMP / pharmacology
  • Cystic Fibrosis / genetics*
  • Cystic Fibrosis / physiopathology
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA / genetics
  • DNA / isolation & purification
  • Gene Expression
  • Humans
  • Ion Channel Gating / drug effects
  • Ion Channels / genetics*
  • Ion Channels / physiology
  • Kinetics
  • Lung / physiopathology
  • Membrane Proteins / genetics*
  • Membrane Proteins / physiology
  • Probability
  • Promoter Regions, Genetic
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • Vaccinia virus / genetics

Substances

  • CFTR protein, human
  • Chloride Channels
  • Ion Channels
  • Membrane Proteins
  • RNA, Messenger
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA
  • Cyclic AMP