Association of degradation and secretion of three chimeric polypeptides in Escherichia coli

R. Gentz, Y. Kuys, Christian W Zwieb, D. Taatjes, H. Taatjes, W. Bannwarth, D. Stueber, I. Ibrahimi

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

We investigated the stability of fusion proteins composed of the signal peptide of the heat-labile enterotoxin of Escherichia coli and three polypeptides: the bacterial cytoplasmic chloramphenicol acetyltransferase, the mouse dihydrofolate reductase, and human immune interferon. We demonstrate that these proteins are rapidly degraded as a result of being targeted to the secretion apparatus of E. coli, with the extent of degradation varying among the three fusion proteins. Four lines of experimental evidence are presented in support of this suggestion. First, the chimeric polypeptides containing a functional signal peptide were detected in low amounts in vivo. When a mutation was introduced in the signal peptide, resulting in lack of recognition by the secretion apparatus, the chimeric proteins accumulated at high levels in the cytoplasma of the cell. Second, both the wild-type and mutant polypeptidase accumulated in a purified and reconstituted in vitro translation system from E. coli and were equally susceptible to digestion by an exogenous protease. Third, the chimeric polypeptides lacking the signal peptide accumulated in a stable form in vivo. Fourth, the precursors of the proteins containing a functional signal peptide accumulated in a secA ts mutant at the restrictive temperature when secretion was blocked, suggesting that degradation is tightly linked to the secretion apparatus.

Original languageEnglish (US)
Pages (from-to)2212-2220
Number of pages9
JournalJournal of Bacteriology
Volume170
Issue number5
StatePublished - 1988
Externally publishedYes

Fingerprint

Protein Sorting Signals
Escherichia coli
Peptides
Tetrahydrofolate Dehydrogenase
Proteins
Chloramphenicol O-Acetyltransferase
Protein Precursors
Protein Stability
Enterotoxins
Interferon-gamma
Digestion
Peptide Hydrolases
Hot Temperature
Mutation
Temperature

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Gentz, R., Kuys, Y., Zwieb, C. W., Taatjes, D., Taatjes, H., Bannwarth, W., ... Ibrahimi, I. (1988). Association of degradation and secretion of three chimeric polypeptides in Escherichia coli. Journal of Bacteriology, 170(5), 2212-2220.

Association of degradation and secretion of three chimeric polypeptides in Escherichia coli. / Gentz, R.; Kuys, Y.; Zwieb, Christian W; Taatjes, D.; Taatjes, H.; Bannwarth, W.; Stueber, D.; Ibrahimi, I.

In: Journal of Bacteriology, Vol. 170, No. 5, 1988, p. 2212-2220.

Research output: Contribution to journalArticle

Gentz, R, Kuys, Y, Zwieb, CW, Taatjes, D, Taatjes, H, Bannwarth, W, Stueber, D & Ibrahimi, I 1988, 'Association of degradation and secretion of three chimeric polypeptides in Escherichia coli', Journal of Bacteriology, vol. 170, no. 5, pp. 2212-2220.
Gentz R, Kuys Y, Zwieb CW, Taatjes D, Taatjes H, Bannwarth W et al. Association of degradation and secretion of three chimeric polypeptides in Escherichia coli. Journal of Bacteriology. 1988;170(5):2212-2220.
Gentz, R. ; Kuys, Y. ; Zwieb, Christian W ; Taatjes, D. ; Taatjes, H. ; Bannwarth, W. ; Stueber, D. ; Ibrahimi, I. / Association of degradation and secretion of three chimeric polypeptides in Escherichia coli. In: Journal of Bacteriology. 1988 ; Vol. 170, No. 5. pp. 2212-2220.
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