[Isolation and certain properties of the dermonecrotic toxin from Pasteurella multocida]. / Vydelenie i nekotorye svoistva dermonekroticheskogo toksina Pasteurella multocida.

The procedure for isolation and purification of Pasteurella multocida serovariant D toxin has been described. It includes the three steps of protein precipitation from cultural filtrates by 70% ammonium sulfate, chromatography of the concentrated material on Ultragel AcA44 gel-filtration on Sephracryl S-200. The proposed technique permits one the 155-fold purification of the preparation with 32.6% yield estimated by biological activity. The obtained purified preparation is homogeneous in polyacrylamide gel electrophoresis. The immunological methods also confirm the homogeneity of the preparation. The minimal dermonecrotic dose for guinea pigs of the purified 120 kDa toxin is 78 ng and LD50 for mice is 280 ng. Pasteurella multocida toxin is found to be a thermolabile protein sensitive to trypsin, glutaraldehyde and formaldehyde treatments.

The characterization of ion channels formed by Pasteurella multocida dermonecrotic toxin.

The influence of the dermonecrotic lethal toxin (approximately 120 kDa) produced by Pasteurella multocida serovarian D on planar phospholipid bilayers was studied. It was found that the toxin is able to increase the conductance of the bilayers by formation of low-conductive and cation-selective ion channels [27 pS at 4.0 M KCl, pH 7.5; zero current potential equals to -14.5 +/- 0.5 mV at threefold transmembrane gradient KCl (120 mM/40 mM)]. In biionic conditions the channels displayed weak selectivity between Na, K and Ca ions. The shapes of current-voltage characteristics (which were measured at different pH and salt concentrations) indicate that an energetic barrier for passing ions is situated near the center of the water pore of the ion channels. The effective diameter of the ion channel's water pore was established to be equal to 2.1 +/- 0.3 nm.