Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1993 May;61(5):2200–2202. doi: 10.1128/iai.61.5.2200-2202.1993

Membrane translocation and channel-forming activities of diphtheria toxin are blocked by replacing isoleucine 364 with lysine.

V Cabiaux 1, J Mindell 1, R J Collier 1
PMCID: PMC280822  PMID: 7683003

Abstract

A mutant of diphtheria toxin in which Ile-364 was replaced by Lys was at least 500-fold less toxic to Vero cells than the parental toxin. Its ability to undergo low-pH-triggered translocation across the plasma membrane was greatly diminished, as was its ability to form ion-conductive channels. In addition, the mutant toxin was inactive in the pH-dependent killing of Escherichia coli.

Full text

PDF
2201

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrams C. K., Jakes K. S., Finkelstein A., Slatin S. L. Identification of a translocated gating charge in a voltage-dependent channel. Colicin E1 channels in planar phospholipid bilayer membranes. J Gen Physiol. 1991 Jul;98(1):77–93. doi: 10.1085/jgp.98.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cabiaux V., Phalipon A., Wattiez R., Falmagne P., Ruysschaert J. M., Kaczorek M. Expression of a biologically active diphtheria toxin fragment B in Escherichia coli. Mol Microbiol. 1988 May;2(3):339–346. doi: 10.1111/j.1365-2958.1988.tb00037.x. [DOI] [PubMed] [Google Scholar]
  3. Carroll S. F., Collier R. J. Diphtheria toxin: quantification and assay. Methods Enzymol. 1988;165:218–225. doi: 10.1016/s0076-6879(88)65034-8. [DOI] [PubMed] [Google Scholar]
  4. Carter P., Bedouelle H., Winter G. Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res. 1985 Jun 25;13(12):4431–4443. doi: 10.1093/nar/13.12.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Choe S., Bennett M. J., Fujii G., Curmi P. M., Kantardjieff K. A., Collier R. J., Eisenberg D. The crystal structure of diphtheria toxin. Nature. 1992 May 21;357(6375):216–222. doi: 10.1038/357216a0. [DOI] [PubMed] [Google Scholar]
  6. Donovan J. J., Simon M. I., Draper R. K., Montal M. Diphtheria toxin forms transmembrane channels in planar lipid bilayers. Proc Natl Acad Sci U S A. 1981 Jan;78(1):172–176. doi: 10.1073/pnas.78.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Falmagne P., Capiau C., Lambotte P., Zanen J., Cabiaux V., Ruysschaert J. M. The complete amino acid sequence of diphtheria toxin fragment B. Correlation with its lipid-binding properties. Biochim Biophys Acta. 1985 Jan 21;827(1):45–50. doi: 10.1016/0167-4838(85)90099-8. [DOI] [PubMed] [Google Scholar]
  8. London E. Diphtheria toxin: membrane interaction and membrane translocation. Biochim Biophys Acta. 1992 Mar 26;1113(1):25–51. doi: 10.1016/0304-4157(92)90033-7. [DOI] [PubMed] [Google Scholar]
  9. Mekada E., Uchida T. Binding properties of diphtheria toxin to cells are altered by mutation in the fragment A domain. J Biol Chem. 1985 Oct 5;260(22):12148–12153. [PubMed] [Google Scholar]
  10. O'Keefe D. O., Cabiaux V., Choe S., Eisenberg D., Collier R. J. pH-dependent insertion of proteins into membranes: B-chain mutation of diphtheria toxin that inhibits membrane translocation, Glu-349----Lys. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6202–6206. doi: 10.1073/pnas.89.13.6202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. O'Keefe D., Collier R. J. Cloned diphtheria toxin within the periplasm of Escherichia coli causes lethal membrane damage at low pH. Proc Natl Acad Sci U S A. 1989 Jan;86(1):343–346. doi: 10.1073/pnas.86.1.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olsnes S., Stenmark H., Moskaug J. O., McGill S., Madshus I. H., Sandvig K. Protein toxins with intracellular targets. Microb Pathog. 1990 Mar;8(3):163–168. doi: 10.1016/0882-4010(90)90043-p. [DOI] [PubMed] [Google Scholar]
  13. Sandvig K., Olsnes S. Rapid entry of nicked diphtheria toxin into cells at low pH. Characterization of the entry process and effects of low pH on the toxin molecule. J Biol Chem. 1981 Sep 10;256(17):9068–9076. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES