Abstract
CYSTIC fibrosis (CF) is a common lethal genetic disease that manifests itself in airway and other epithelial cells as defective chloride ion absorption and secretion1,2, resulting at least in part from a defect in a cyclic AMP-regulated, outwardly-rectifying Cl− channel in the apical surface3–5. The gene responsible for CF has been identified and predicted to encode a membrane protein termed the CF transmembrane conductance regulator (CFTR)6–8. Identification of a cryptic bacterial promoter within the CFTR coding sequence led us to construct a complementary DNA in a low-copy-number plasmid, thereby avoiding the deleterious effects of CFTR expression on Escherischia coli. We have used this cDNA to express CFTR in vitro and in vivo. Here we demonstrate that CFTR is a membrane-associated glycoprotein that can be phosporylated in vitro by cAMP-dependent protein kinase. Polyclonal and monoclonal antibodies directed against distinct domains of the protein immunoprecipitated recombinant CFTR as well as the endogenous CFTR in nonrecombinant T84 cells. Partial proteolysis fingerprinting showed that the recombinant and non-recombinant proteins are indistinguishable. These data, which establish several characteristics of the protein responsible for CF, will now enable CFTR function to be studied and will provide a basis for diagnosis and therapy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Boat, T., Welsh, M. & Beaudet, A. in The Metabolic Basis of Inherited Disease (eds Scriver C, Beaudet A., Sly, W. & Valle, D.) 2649–2680 (McGraw Hill, New York, 1989).
Quinton, P. Clin. Chem. 35, 726–730 (1989).
Li, M. et al. Nature 331, 358–360 (1988).
Frizzell, R., Rechkemmer, G. & Shoemaker, R. Science 233, 558–560 (1986).
Welsh, M. Science 232, 1648–1650 (1986).
Rommens, J. et al., Science 245, 1059–1065 (1989).
Riordan, J. et al., Science 245, 1066–1073 (1989).
Kerem, B.-S. et al. Science 245, 1073–1080 (1989).
Brosius, J. Gene 27, 151–160 (1984).
Hawley, D. K. & McClure, W. R. Nucleic Acids Res. 11, 2237–2255 (1983).
Cohen, S. N., Chang, A. C. Y., Boyer, H. W. & Heling, R. B. Proc. natn. Acad. Sci. U.S.A. 70, 3240–3244 (1973).
Jang, S. K., Davies, M. V., Kaufman, R. J., & Wimmer, E. J. Virol. 63, 1651–1660 (1989).
Elroy-Stein, O., Fuerst, T. R. & Moss, B. Proc. natn. Acad. Sci. U.S.A. 86, 6126–6130 (1989).
Greenberger, L. M., Williams, S. S. & Horowitz, S. B. J. biol. Chem. 262, 13685–13689 (1987).
Nash, B. & Tate, S. S. J. biol. Chem. 259, 678–684 (1984).
Richard, N. D., Aldwin, L., Nitecki, D., Gottesman, M. & Pastan, I. Biochemistry 27, 7607–7613 (1988).
Fuerst, T. R., Niles, E. G., Studier, W. & Moss, B. Proc. natn. Acad. Sci. U.S.A. 83, 8122–8126 (1986).
Kohler, G. & Milstein, C. Nature 256, 495–497 (1975).
Cheng, S. H. et al. EMBO J. 7, 3845–3855 (1988).
Harlow, E., Crawford, L. V., Pim, D. C. & Williamson, N. M. J. Virol. 39, 861–869 (1981).
Wurzner, R., Oppermann, M., Zierz, R., Baumgarten, H. & Gotze, O. J. Immun. Meths. 126, 231–237 (1980).
Kawata, M. et al. J. biol. Chem. 264, 15688–15695 (1989).
Muriakarmi, H. & Masui, H. Proc. natn. Acad. Sci. U.S.A. 77, 3464–3468 (1980).
Lieber, M., Mazetta, J., Nelson-Rees, W., Kaplan, M. & Todaro, G. Int. J. Cancer 15, 741–747 (1975).
Rich, D. P. et al. Nature 347, 358–363 (1990).
Sambrook, J., Fritsch, E. F. & Maniatis, T. in Molecular Cloning, A Laboratory Manual (Cold Spring Harbor Laboratory, New York, 1989).
Stoker, N. G., Fairweather, N. F. & Spratt, B. G. Gene 18, 335–341 (1982).
Laemmli, U. K. Nature 227, 680–685 (1970).
Tarentino, A. L., Gomez, C. M. & Plummer, T. H. Biochemistry 24, 4665–4671 (1985).
Feigner, P. L. et al. Proc. natn. Acad. Sci. U.S.A. 84, 7413–7417 (1987).
Harlow, E. & Lane, D. In Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory, New York, 1988).
Harvey, R., Hehir, K. M., Smith, A. E. & Cheng, S. H. Molec. cell. Biol. 9, 3647–3656 (1989).
Mole, S. & Lane, D. J. Virol. 54, 703–710 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gregory, R., Cheng, S., Rich, D. et al. Expression and characterization of the cystic fibrosis transmembrane conductance regulator. Nature 347, 382–386 (1990). https://doi.org/10.1038/347382a0
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1038/347382a0
This article is cited by
-
Impact of gene editing on the study of cystic fibrosis
Human Genetics (2016)
-
Glucose-induced electrical activities and insulin secretion in pancreatic islet β-cells are modulated by CFTR
Nature Communications (2014)
-
Intrinsic predisposition of naïve cystic fibrosis T cells to differentiate towards a Th17 phenotype
Respiratory Research (2013)
-
Proteases, cystic fibrosis and the epithelial sodium channel (ENaC)
Cell and Tissue Research (2013)
-
Initial interrogation, confirmation and fine mapping of modifying genes: STAT3, IL1B and IFNGR1 determine cystic fibrosis disease manifestation
European Journal of Human Genetics (2011)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
