Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae

K. U. Jansen, W. G. Conroy, T. Claudio, T. D. Fox, N. Fujita, Owen Hamill, J. M. Lindstrom, M. Luther, N. Nelson, K. A. Ryan, M. T. Sweet, G. P. Hess

Research output: Contribution to journalArticle

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Abstract

Yeast expression vectors were constructed containing complementary DNA encoding the α-, β-, γ-, and δ-subunits of the Torpedo californica nicotinic acetylcholine receptor under the control of the Saccharomyces cerevisiae alcohol dehydrogenase promoter. All four plasmids were integrated into the yeast genome of a single yeast cell. The resulting yeast strain synthesized polypeptides novel to yeast that had the molecular weights and antigenic properties similar to the authentic T. californica receptor α-, γ-, and δ-subunits. The β-subunit polypeptide could not be detected in this yeast strain, even though the poly(A)+ RNA from this strain contained all the information necessary for the expression of functional acetylcholine receptors in Xenopus laevis oocytes. The replacement of the β-subunit mRNA 5'-untranslated leader and its N-terminal signal sequence by the corresponding α-subunit sequences, however, resulted in the expression of the β-subunit polypeptide in yeast grown at 5°C.

Original languageEnglish (US)
Pages (from-to)15022-15027
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number25
StatePublished - 1989
Externally publishedYes

Fingerprint

Torpedo
Nicotinic Receptors
Yeast
Saccharomyces cerevisiae
Yeasts
Peptides
Messenger RNA
Alcohol Dehydrogenase
Xenopus laevis
Cholinergic Receptors
Protein Sorting Signals
Oocytes
Plasmids
Complementary DNA
Molecular Weight
Genome
Genes
Molecular weight
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Jansen, K. U., Conroy, W. G., Claudio, T., Fox, T. D., Fujita, N., Hamill, O., ... Hess, G. P. (1989). Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae. Journal of Biological Chemistry, 264(25), 15022-15027.

Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae. / Jansen, K. U.; Conroy, W. G.; Claudio, T.; Fox, T. D.; Fujita, N.; Hamill, Owen; Lindstrom, J. M.; Luther, M.; Nelson, N.; Ryan, K. A.; Sweet, M. T.; Hess, G. P.

In: Journal of Biological Chemistry, Vol. 264, No. 25, 1989, p. 15022-15027.

Research output: Contribution to journalArticle

Jansen, KU, Conroy, WG, Claudio, T, Fox, TD, Fujita, N, Hamill, O, Lindstrom, JM, Luther, M, Nelson, N, Ryan, KA, Sweet, MT & Hess, GP 1989, 'Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae', Journal of Biological Chemistry, vol. 264, no. 25, pp. 15022-15027.
Jansen, K. U. ; Conroy, W. G. ; Claudio, T. ; Fox, T. D. ; Fujita, N. ; Hamill, Owen ; Lindstrom, J. M. ; Luther, M. ; Nelson, N. ; Ryan, K. A. ; Sweet, M. T. ; Hess, G. P. / Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 25. pp. 15022-15027.
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