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 Huganir; Peter Maloney; William B. Guggino

doi:10.1073/pnas.89.1.344

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 Huganir, Peter Maloney, William B. Guggino

School of Medicine

Research output: Contribution to journal › Article › peer-review

62 Scopus citations

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 language	English (US)
Pages (from-to)	344-347
Number of pages	4
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	89
Issue number	1
DOIs	https://doi.org/10.1073/pnas.89.1.344
State	Published - 1992

Keywords

Chloride channel
Cystic fibrosis transmembrane conductance regulator

ASJC Scopus subject areas

General

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10.1073/pnas.89.1.344

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Zeitlin, P. L., Crawford, I., Lu, L., Woel, S., Cohen, M. E., Donowitz, M., Montrose, M. H., Hamosh, A., Cutting, G. R., Gruenert, D., Huganir, R., Maloney, P., & Guggino, W. B. (1992). CFTR protein expression in primary and cultured epithelia. Proceedings of the National Academy of Sciences of the United States of America, 89(1), 344-347. https://doi.org/10.1073/pnas.89.1.344

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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.",

keywords = "Chloride channel, Cystic fibrosis transmembrane conductance regulator",

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

year = "1992",

doi = "10.1073/pnas.89.1.344",

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T1 - CFTR protein expression in primary and cultured epithelia

AU - Zeitlin, Pamela L.

AU - Crawford, Isabelle

AU - Lu, Luo

AU - Woel, Sybille

AU - Cohen, Michael E.

AU - Donowitz, Mark

AU - Montrose, Marshall H.

AU - Hamosh, Ada

AU - Cutting, Garry R.

AU - Gruenert, Dieter

AU - Huganir, Richard

AU - Maloney, Peter

AU - Guggino, William B.

PY - 1992

Y1 - 1992

N2 - 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.

AB - 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.

KW - Chloride channel

KW - Cystic fibrosis transmembrane conductance regulator

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CFTR protein expression in primary and cultured epithelia

Abstract

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