Biomedical subjects
Publications and source records attributed to E I Grebinozhko.
A simple and relatively rapid method is elaborated for development of electrophoregram in PAAG by silver binding directly with protein. This permits avoiding the gel staining, considerably improves the quality of staining and promotes better reproducibility of the results. Sensitivity of the method permits detecting 1 ng of protein in the band.
Interaction of latrotoxin with phospholipid vesicles and bilayer lipid membranes is shown to proceed differently. Latrotoxin when interacting with liposomes is sorbed on the membrane surface forming no ionic channels in this case. Only latrotoxin fragments obtained due to the toxin hydrolysis by pronase or trypsin are able to form channels. These fragments being inserted into liposomes are coupled strongly with the membrane and are not subjected to the further splitting by proteinases. The electrophoretic spectrum of peptides bound with liposomes is presented by protein components with a molecular weight of 116, 100, 92, 67, 52 and 45 kDa, while zone typical of latrotoxin is absent in this spectrum. The method of small-angle X-rays scattering has shown that tryptic fragments of latrotoxin penetrate into the phospholipid bilayer of liposomes.
alpha-Latrotoxin, a main toxic component of the Latrodectus mactans tredecimguttatus venom is a large polypeptide with molecular weight of 130 KDa. A rapid method is suggested for isolating this protein using high-effective liquid chromatography on chromatograph FPLC ("Pharmacia", Sweden). The isolated protein does not differ from the previously described alpha-latrotoxin in the main physicochemical parameters as well as in physiological properties.