CFTR protein expression in primary and cultured epithelia

Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):344-7. doi: 10.1073/pnas.89.1.344.

Abstract

The gene responsible for the lethal disorder cystic fibrosis encodes a 1480-amino acid glycoprotein, CFTR. Using polyclonal antibodies directed against separate phosphorylation sites in the pre-nucleotide-binding fold (exon 9) and the R domain (exon 13), we have identified a 165-kDa protein in Xenopus laevis oocytes injected with recombinant CFTR cRNA transcribed from the full-length CFTR plasmid pBQ4.7. A protein of the same mobility was also detected with Western blotting techniques in whole cell extracts of cells that express CFTR mRNA (T84, FHTE, HT-29), including biopsied human nasal and bronchial tissue. Immunodetectable 165-kDa protein was concentrated in the apical membrane fraction of ileal villus tissue. We also report that the 165-kDa protein levels can be modulated pharmacologically, and these levels are appropriately correlated with second-messenger-regulated Cl- efflux. Thus, native or recombinant CFTR can be recognized by these anti-CFTR peptide polyclonal antibodies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Cell Polarity
  • Cells, Cultured
  • Chloride Channels
  • Cystic Fibrosis / metabolism*
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Epithelium / metabolism*
  • Humans
  • In Vitro Techniques
  • Membrane Proteins / immunology
  • Membrane Proteins / metabolism*
  • Oocytes
  • Peptides / immunology
  • Rabbits
  • Xenopus laevis

Substances

  • CFTR protein, human
  • Chloride Channels
  • Membrane Proteins
  • Peptides
  • Cystic Fibrosis Transmembrane Conductance Regulator