Biomedical subjects

O V Krasilnikov

Publications and source records attributed to O V Krasilnikov.

27 records · Page 2Linked to original sources

PubMed1993

Relation between ionic channel conductance and conductivity of media containing different nonelectrolytes. A novel method of pore size determination.

The effects of nonelectrolytes on conductivity and viscosity of KCl solutions as well as on ion channel conductance were studied. Mobility of ions in solutions were found to solely depend on percent concentration (w/w) of the nonelectrolytes added and to be effectively independent on their chemical nature (sugars or polyglycols) and molecular size. Proportional changes in both the ion channel conductance and the conductivity of bulk solution induced by low m. w. nonelectrolytes may be used as a criterion of diffusion mechanism of ion transport through channels. The slope of the dependence of ion channel conductance on conductivity of bulk solution containing different concentrations of nonelectrolytes is a good measure of channel permeability for nonelectrolyte. A new method of pore size determination is introduced. Results of practical application of this simple method to three types of ion channels (formed by alpha-latrotoxin, staphylococcal alpha-toxin and its N-terminal fragment) are shown. The advantages and disadvantages of the method are discussed.

Bacterial Toxins↗

PubMed1993

The interaction of amphotericin B with cell membrane of rat thymocytes.

Amphotericin B (AB) at micromolar concentrations increases cell membrane permeability and induced swelling of rat thymus lymphocytes. Potassium efflux is a precondition for AB to induce swelling of the cells. The rate constants for potassium loss and volume changes were proportional to the 1.24th and the 2nd power of the antibiotic concentration respectively. The reflection coefficients for nonelectrolytes with different hydrodynamic radii were determined, and the equivalent radius of the amphotericin pore in the thymocyte cell membrane was estimated to be 4.1 +/- 0.3 A at polyene concentrations varying between 2.5 mumol/l and 80 mumol/l. It is suggested that channel formation by AB in cell membranes is actually able to modulate immune responses.

Amphotericin B↗

PubMed1992

Comparative analysis of latrotoxin channels of different conductance in planar lipid bilayers. Evidence for cluster organization.

It has been established that channels induced by Latrodectus tredicimguttatus alpha-toxin (LT) in lipid bilayers have a cluster organisation. So far as: (i) the LT-channels had practically identical sizes of its water pores (r = 9.4 +/- 0.6 A) independently on the lipid composition of planar bilayer lipid membrane (BLM) although their conductances might differ from each other more than 10 times (100 mM KCl (pH 7.5)). (ii) affinity of permeable ions to channels had a small variation with distinct group of BLM, although LT-channels conductances varied from 112 +/- 8 pS till 1110 +/- 40 pS for phosphatidylcholine-BLM and from 75 +/- 6 pS till 170 +/- 14 pS for phosphatidylserine-BLM. (iii) Ca/K selectivity was greater in negatively charged membranes but did not also depend on the channel amplitude for the same BLM. Cation-anionic selectivity was identical for all studied channels.

Lipid Bilayers↗

PubMed1992

The mode of action of Vibrio cholerae cytolysin. The influences on both erythrocytes and planar lipid bilayers.

The interaction with erythrocytes of cholera cytolysin (CC) obtained from a non-01 Vibrio cholerae strain results in the osmotic rupture of target cells upon formation by CC of the waterfilled pores in their membranes. The aggregation of several toxin monomers is required for the formation of one CC channel with a radius of 0.9-1.0 nm. The investigations using planar bilayer lipid membranes suggest that the CC-induced pore is an interprotein anion selective channel carrying a fixed positive charge. The role of the charge was supported by the influence of pH on the selectivity, single conductance and voltage gating of the CC channels. The ability of the CC to modify both model and natural membranes has a maximum at pH 6.0-7.0. It was found that CC channels insert into the membrane asymmetrically. The effect of proteolytic treatment of the channel by papain also indicates that the two entrances of the channel protrude from the plane of the membrane into the solution for different distances. It is proposed that the biological effects of the non-01 V. cholera cytolysin are based on its channel-forming activity.

Animals↗

PubMed1992

A simple method for the determination of the pore radius of ion channels in planar lipid bilayer membranes.

A new method of pore size determination is presented. The results of applying this simple method to ion channels formed by staphylococcal alpha-toxin and its N-terminal fragment as well as to cholera toxin channels are shown. The advantages and the difficulties of this method are discussed. It was found that (i) the mobility of ions in solutions depends only on the percentage of concentration of added non-electrolytes and practically not on their chemical nature (sugars or polyglycols) and molecular size; (ii) the proportional change of both ion channel conductance and bulk solution conductivity by low M. nonelectrolytes may be used as an indication of a diffusion mechanism of ion transport through channels; (iii) the slope of the dependence of the ion channel conductance on the bulk conductivity of solutions containing different concentrations of non-electrolyte is a good measure of channel permeability for non-electrolytes.

Bacterial Toxins↗

PubMed1991

The ionic channels formed by cholera toxin in planar bilayer lipid membranes are entirely attributable to its B-subunit.

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.

Anions↗

PubMed1990

Memory is a property of an ion channels pool: ion channels formed by Staphylococcus aureus alpha-toxin.

The short-time depolarization effects on the integral conductance induced by S. aureus alpha-toxin (ST) in planar lipid bilayer membranes has been studied. Ion channels formed by ST were found to have several potential-induced nonconductance (closed) states. The transitions of ion channels between the states are only through one conductance state. The transition of ST-channels from closed to open state is induced by membrane depolarization. The amplitude current after a series of voltage pulses is a function of pulse number, and is effectively independent of the time interval between the neighbouring pulses. Therefore, a membrane which contains a pool of ion channels "remembers" its previous existence. A simple model can be used to explain this phenomenon.

Bacterial Toxins↗

PubMed1989

Ion transport through channels formed in lipid bilayers by Staphylococcus aureus alpha-toxin.

Staphylotoxin channel appears to be predominantly anion-selective with non-linear and asymmetric current-voltage characteristics (CVC) at neutral pH. Increased salt concentrations induce linearity and asymmetry of CVC and loss of selectivity. At lower pH both the channel conductivity and anion selectivity increase. Higher temperatures raise the channel conductivity in parallel with the changes in electrical conductivity of the salt solution, but do not change selectivity. Experimental dependences are described obtained by approximation of electrical diffusion and considering the interactions of penetrating ions with fixed charges at the entrances and the channel energy profile.

Bacterial Toxins↗

PubMed1988

The structure of Staphylococcus aureus alpha-toxin-induced ionic channel.

Polyethylene glycols (PEG) with molecular weight less than or equal to 3000 were shown to effectively protect human erythrocytes from osmotic lysis induced by alpha-staphylotoxin (ST). PEG with MW less than 3000 do not change the conductivity of ion channels induced by ST in bilayer lipid membranes (BLM). Changing the bilayer from a pure phosphatidylcholine (PC) to a negatively charged phosphatidylserine (PS) film results in an asymmetry of the current-voltage characteristics. This is evidenced by the asymmetrical position of the ST-channel pore in bilayer membranes. The results obtained allow to conclude that the ST-channel is an interprotein pore filled with water (with an inner diameter of 2.5-3 nm and a length of approximately 10 nm). It is composed of six molecules of alpha-toxin from Staphylococcus aureus. The ST-channel incorporates into a membrane with only one mouth in contact with the polar lipid heads and the other one protruding 4.5-5 nm from the bilayer plane in water solution.

Bacterial Toxins↗