Membrane topology of the colicin A pore-forming domain analyzed by disulfide bond engineering

Denis Duché, Jacques Izard, Juan M. González-Mañas, Michael W. Parker, Marcel Crest, Martine Chartier, Daniel Baty

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Four colicin A double-cysteine mutants possessing a disulfide bond in their pore-forming domain were constructed to study the translocation and the pore formation of colicin A. The disulfide bonds connected α-helices 1 and 2, 2 and 10, 3 and 9, or 3 and 10 of the pore-forming domain. The disulfide bonds did not prevent the colicin A translocation through the Escherichia coli envelope. However, the mutated colicins were able to exert their in vivo channel activity only after reduction of their disulfide bonds. In vitro studies with brominated phospholipid vesicles and planar lipid bilayers revealed that the disulfide bond that connects the α-helices 2 and 10 prevented the colicin A membrane insertion, whereas the other double-cysteine mutants inserted into lipid vesicles. The disulfide bonds that connect either the α-helices 1 and 2 or 3 and 10 were unable to prevent the formation of a conducting channel in presence of membrane potential. These results indicate that α-helices 1, 2, 3, and 10 remain at the membrane surface after application of a membrane potential.

Original languageEnglish (US)
Pages (from-to)15401-15406
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number26
DOIs
StatePublished - Jul 16 1996

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Colicins
Disulfides
Topology
Membranes
Membrane Potentials
Cysteine
Lipid bilayers
Lipid Bilayers
Escherichia coli
Phospholipids
Lipids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Duché, D., Izard, J., González-Mañas, J. M., Parker, M. W., Crest, M., Chartier, M., & Baty, D. (1996). Membrane topology of the colicin A pore-forming domain analyzed by disulfide bond engineering. Journal of Biological Chemistry, 271(26), 15401-15406. https://doi.org/10.1074/jbc.271.26.15401

Membrane topology of the colicin A pore-forming domain analyzed by disulfide bond engineering. / Duché, Denis; Izard, Jacques; González-Mañas, Juan M.; Parker, Michael W.; Crest, Marcel; Chartier, Martine; Baty, Daniel.

In: Journal of Biological Chemistry, Vol. 271, No. 26, 16.07.1996, p. 15401-15406.

Research output: Contribution to journalArticle

Duché, D, Izard, J, González-Mañas, JM, Parker, MW, Crest, M, Chartier, M & Baty, D 1996, 'Membrane topology of the colicin A pore-forming domain analyzed by disulfide bond engineering', Journal of Biological Chemistry, vol. 271, no. 26, pp. 15401-15406. https://doi.org/10.1074/jbc.271.26.15401
Duché, Denis ; Izard, Jacques ; González-Mañas, Juan M. ; Parker, Michael W. ; Crest, Marcel ; Chartier, Martine ; Baty, Daniel. / Membrane topology of the colicin A pore-forming domain analyzed by disulfide bond engineering. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 26. pp. 15401-15406.
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