Lipid interaction of diphtheria toxin and mutants with altered fragment B. 1. Liposome aggregation and fusion.
The interaction of diphtheria toxin and its cross-reacting mutants crm 45,228 and 1001 with small unilamellar vesicles has been followed by a turbidity assay, electron microscopy, fluorescence energy transfer and membrane permeability. All toxins at pH lower than 6 induce the aggregation and fusion of liposomes containing negatively charged phospholipids; crm 45 and crm 1001 are less potent than diphtheria toxin. Isolated diphtheria toxin fragment B is very effective while isolated fragment A is ineffective. Liposome fusion induced by the toxins at low pH occurs without release of the internal content implying that fusion does not involve vesicle breakage and resealing. The pH dependence of the membrane interaction of diphtheria toxin monitored by turbidity is in close agreement with that monitored by fluorescence energy transfer. It shows that diphtheria toxin can alter the lipid bilayer structure in the pH interval 5-6. This pH range occurs in endosomes and suggests that histidyl and carboxyl residues are likely to be involved in the conformational change of diphtheria toxin triggered by acidic pH.
Papini
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E
Centro Consiglio Nazionale delle Ricerche per le Biomembrane, Università di Padova, Italy.
Colonna
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R
Cusinato
F
F
Montecucco
C
C
Tomasi
M
M
Rappuoli
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D004167:Diphtheria Toxin / Q000235:genetics / Q000378:metabolism*; D004735:Energy Transfer; D006863:Hydrogen-Ion Concentration; D050356:Lipid Metabolism; D008081:Liposomes / Q000378:metabolism*; D008854:Microscopy, Electron; D009154:Mutation; D009391:Nephelometry and Turbidimetry; D010446:Peptide Fragments / Q000378:metabolism; D010539:Permeability
D004167:Diphtheria Toxin; D008081:Liposomes; D010446:Peptide Fragments; C039171:diphtheria toxin fragment B