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1.
Biochim Biophys Acta ; 1067(2): 166-70, 1991 Aug 26.
Article in English | MEDLINE | ID: mdl-1715187

ABSTRACT

The interaction of cholera toxin with planar bilayer lipid membranes (BLM) at low pH results in the formation of ionic channels, the conductance of which can be directly measured in voltage-clamp experiments. It is found that the B-subunit of cholera toxin (CT-B) also is able to induce ionic channels in BLM whereas the A-subunit is not able to do it. The increase of pH inhibited the channel-forming activity of CT-B. The investigation of pH-dependences of both the conductance and the cation-anion selectivity of the CT-B channel allowed us to suggest that the water pore of this channel is confined to the B-subunit of cholera toxin. The effective diameter of the CT-B channels water pores was directly measured in BLM and is equal to 2.1 +/- 0.2 nm. The channels formed by whole toxin and its B-subunit exhibit voltage-dependent activity. We believe these channels are relevant to the mode of action of cholera toxin and especially to the endosomal pathway of the A-subunit into cells.


Subject(s)
Cholera Toxin/metabolism , Ion Channels/physiology , Lipid Bilayers , Anions , Cations , Hydrogen-Ion Concentration , Membrane Potentials
2.
Biull Eksp Biol Med ; 104(10): 472-5, 1987 Oct.
Article in Russian | MEDLINE | ID: mdl-2445393

ABSTRACT

Antigenic determinants of subunits A and B of cholera enterotoxin (CT), heat-labile enterotoxin from the human E. coli strain (hLT) and heat-labile enterotoxin from the porcine E. coli strain (pLT) were analysed by Ouchterlony double gel-diffusion test against antisera to B subunits of three toxins and antisera to three holotoxins. The results have shown the existence of the following antigenic determinants: in subunits B-1. antigenic determinants, common for B subunits of all three enterotoxins-B(chp); 2. group antigenic determinants, common for B subunits of two toxins in the pair-B(ch), B(hp); 3. antigenic determinants, unique for B subunits of each CT, hLT, pLT-(B(c), B(h), B(p); in subunits A.-1. antigenic determinants, common for A subunits of all three enterotoxins-A.(chp); 2. group antigenic determinants, common for A subunits of two enterotoxins (hLT and pLT/-A(hp); 3. antigenic determinants, unique for A subunit of CT-A(c). On the basis of these results antigenic formulas for subunits of CT, hLT, pLT were proposed.


Subject(s)
Bacterial Toxins/immunology , Cholera Toxin/immunology , Enterotoxins/immunology , Escherichia coli Proteins , Escherichia coli , Peptide Fragments/immunology , Animals , Antigens, Bacterial/analysis , Bacterial Toxins/analysis , Bacterial Toxins/isolation & purification , Cholera Toxin/analysis , Cholera Toxin/isolation & purification , Electrophoresis, Polyacrylamide Gel , Enterotoxins/analysis , Enterotoxins/isolation & purification , Epitopes/analysis , Escherichia coli/isolation & purification , Humans , Immunochemistry , Immunodiffusion , Molecular Weight , Peptide Fragments/analysis , Peptide Fragments/isolation & purification , Swine/microbiology
3.
Biull Eksp Biol Med ; 103(1): 80-3, 1987 Jan.
Article in Russian | MEDLINE | ID: mdl-3099858

ABSTRACT

The preparation and identification of B subunit of thermolabile enterotoxin produced by A-B+ gene-containing strain are described. The E. coli strain studied is shown to produce protein identical in its molecular properties and antigenic specificity to B subunit obtained from the whole thermolabile enterotoxin. Partial antigenic affinity between B subunits of thermolabile enterotoxin obtained from different sources and B subunit from cholera enterotoxin has been established in immunochemical studies. Electrophoretic and immunochemical analysis has confirmed the absence of A-subunit admixtures in B-subunit preparation obtained from /A-B+/E. coli strain.


Subject(s)
Enterotoxins/analysis , Escherichia coli , Animals , Antigens, Bacterial/analysis , Bacterial Proteins/analysis , Cholera Toxin/analysis , Cholera Toxin/immunology , Cholera Toxin/isolation & purification , Chromatography, Gel , Electrophoresis, Disc , Enterotoxins/immunology , Enterotoxins/isolation & purification , Escherichia coli/immunology , Immunization , Immunodiffusion , Rabbits
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