Biomedical subjects

V Cabiaux

Publications and source records attributed to V Cabiaux.

45 records · Page 3Linked to original sources

PubMed1988

Expression of a biologically active diphtheria toxin fragment B in Escherichia coli.

The toxB gene of Corynebacterium diphtheriae bacteriophage beta encoding the B fragment of diphtheria toxin was cloned into an inducible expression vector. When expressed in Escherichia coli, fragment B was not proteolysed and was indistinguishable, by immunological criteria, from wild-type C. diphtheriae-derived fragment B. Soluble fragment B was partially purified from the cytoplasm by saline precipitation steps and was shown to compete with the wild-type diphtheria toxin for binding to receptors of sensitive eukaryotic cells. A complete diphtheria toxin was reconstituted by formation of the disulphide bridge between purified fragment A and recombinant fragment B, which migrates at the expected Mr on Western blots and which was able to block protein synthesis by ADP-ribosylation of elongation factor-2, thereby indicating that the recombinant fragment B had retained its biological activity.

Animals↗

PubMed1986

Fusion of Newcastle disease virus with liposomes: role of the lipid composition of liposomes.

We demonstrate here that fusion occurs between the membrane of the Newcastle disease virus (NDV) and liposomes. Fluorescence dequenching studies (using Rhodamine-bearing viral envelopes) revealed the mixing of the lipids constituting the viral and liposomal membrane. The digestion of internal viral proteins by trypsin-containing liposomes indicated the mixing of the internal aqueous compartments. This last assay is independent of exchange of lipids between liposomal and viral membrane in the absence of fusion. Investigation of the effects of liposomal composition indicated that the presence of phosphatidylethanolamine and gangliosides are essential to optimize fusion. The fact that the Newcastle disease virus membrane can fuse with liposome also confirms that fusion must be determined by the viral proteins and could be mostly independent of the nature or presence of the host proteins.

Cholesterol↗

PubMed1986

pH dependent insertion of a diphtheria toxin B fragment peptide into the lipid membrane: a conformational analysis.

A peptide of diphtheria toxin B fragment (residues 147-266) has been shown to induce pore formation in lipid bilayer membranes at low pH. Such an effect was obtained at a much lower extent or not at all at pH = 7. The region localized between residues 225 and 246 is highly hydrophobic (27.3% polarity) and characterized by a high concentration of proline residues. Since proline cis-trans isomerization is highly sensitive to the pH of the medium, we investigated the capability of the cis and trans isomers to penetrate into the lipid matrix. Obviously, the cis-trans isomerization of proline 242 and 245, assumed to be imposed by a low pH, uncovers the hydrophobic region and induces its insertion into a lipid layer of dipalmitoylphosphatidylcholine. The lipid matrix destabilization resulting from this process could promote the penetration into the lipid bilayer of an amphipatic structure (153-178) similar to the transverse lipid associating domains of membrane proteins.

Diphtheria Toxin↗

PubMed1985

Tetanus toxin induces fusion and aggregation of lipid vesicles containing phosphatidylinositol at low pH.

We report here on the ability of tetanus toxin to induce, at low pH, fusion and aggregation of lipid vesicles containing phosphatidylinositol. It has been shown that diphtheria toxin is internalized in acidic vacuoles (endosomes) and that the low endosomal pH could induce a protein conformational change responsible for the interaction with the endosomal membranes and the toxin translocation into the cytoplasm. The data here reported indicate that tetanus toxin might interact with lipid membrane in a similar way as diphtheria toxin suggesting for the two proteins an identical mechanism of entry into cells.

Energy Transfer↗

PubMed1985

Structural analogies between protein kinase C activators.

Phorbol esters and diacylglycerols activate protein kinase C but specific structural parameters appear to be required for the enzyme activation. We have analyzed the conformation of potent and not potent diacylglycerols and phorbol esters. The orientation of the CH20H group at C3 of 1,2 diolein is remarkably similar to that of the same group at C-20 of 4 beta phorbol didecanoate and crucial for potency in activating the enzyme. Our data suggest that the new conformational approach here described could be used to rationally design specific inhibitors preventing the effects of tumor promoters and to predict the structure of potential tumor promoters.

Acetylation↗

PubMed1985

The complete amino acid sequence of diphtheria toxin fragment B. Correlation with its lipid-binding properties.

The complete amino acid sequence of fragment B from diphtheria toxin has been determined. The polypeptide chain was split with cyanogen bromide, o-iodosobenzoic acid, clostripain and trypsin; all amino acid sequence analyses were made by automated Edman degradation. Fragment B, which corresponds to the carboxy terminus of the toxin molecule, contains 342 amino acids and has an Mr of 37240. The proposed amino acid sequence fully confirms the structure recently deduced from the nucleotide sequence of the structural gene. The complete sequence is analyzed in relationship with the role of fragment B in the transfer of diphtheria toxin fragment A from the extracellular medium into the cell cytoplasm.

Amino Acid Sequence↗

PubMed1985

Aggregation and fusion of lipid vesicles induced by diphtheria toxin at low pH: possible involvement of the P site and the NAD+ binding site.

Model membranes have been used to study the interaction between diphtheria toxin and lipids. We report here on the ability of this toxin to induce, at low pH, fusion and aggregation of asolectin lipid vesicles. Resonance energy transfer experiments using lipid fluorescent probes make it possible to discriminate between these two processes.

Adenosine Triphosphate↗

PubMed1984

Diphtheria toxin induces fusion of small unilamellar vesicles at low pH.

Model membrane systems such as phospholipid vesicles have been extensively used to study the mechanism of membrane fusion at the molecular level. We report here on the capacity of diphtheria toxin to induce fusion of small unilamellar vesicles of dipalmitoylphosphatidylcholine at low pH. Fluorescence polarization and differential scanning calorimetry make it possible to demonstrate the mixing of the lipid phase. Mixing of the internal aqueous compartments of liposome was established using the terbium fluorescence technique. The analogy of structure and properties between melittin and a diphtheria toxin fragment is discussed.

Calorimetry↗

PubMed1983

Localization in diphtheria toxin fragment B of a region that induces pore formation in planar lipid bilayers at low pH.

Like diphtheria toxin and the N-terminal (Mr 23 000) region of fragment B, CB1 (Mr 13 000), the cyanogen bromide peptide located in the middle region of fragment B is able to induce pore formation in lipid bilayer membrane at low pH. These two peptides (Mr 23 000 and 13 000) share a common segment (Mr 6300) containing the predicted amphipathic, alpha-helical, transverse lipid-associating domain (Mr 2750) of fragment B [J. Cell Biol. (1980) 87, 837-840]. Therefore, we postulated this domain to be responsible for the pore formation ability of diphtheria toxin [Proc. Natl. Acad. Sci. USA (1981) 78, 172-176]. A relationship between the pH dependency of pore formation and the presence of a cluster of prolines in the C-terminal region of CB1 is proposed.

Diphtheria Toxin↗