CFTR protein expression in primary and cultured epithelia

Pamela L. Zeitlin, Isabelle Crawford, Luo Lu, Sybille Woel, Michael E. Cohen, Mark Donowitz, Marshall H. Montrose, Ada Hamosh, Garry R Cutting, Dieter Gruenert, Richard L Huganir, Peter Maloney, William B Guggino

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)344-347
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number1
StatePublished - 1992

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Keywords

  • Chloride channel
  • Cystic fibrosis transmembrane conductance regulator

ASJC Scopus subject areas

  • Genetics
  • General

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