The lectin of the bark of European spindle (Euonymus europaeus L.): purification, physico-chemical characteristics and application in histochemical studies

Abstract

The purification of lectins from the bark and seeds of E. europaeus L. assessment of comparative characteristics of their carbohydrate and histochemical specificity, evaluation of the feasibility of replacing seed lectin with lectin of the bark of E. europaeus L. Methods. Salting out of NaCl lectins, ion-exchange chromatography on DEAE-Toyopearl and affinity chromatography on cross-linked ovomucin, electrophoretic purity studies in PAAG, comparative study of the immunochemical and histochemical properties of bark and seed lectins. Results. A lectin found in the bark of the European spindle does not differ in carbohydrate specificity from seed lectin. A method has been developed for purifying the found lectin by salting out NaCl, ion-exchange chromatography on DEAE-Toyopearl and affinity chromatography on cross-linked ovomucin by elution with borate buffer solution (pH 9.0 -9.8) in the temperature range +30 -+ 55°C. Electrophoretic purity for lectin from bark was studied and the immunochemical properties of the lectins from bark and seeds were compared. A histochemical study of the binding of bark lectin and European spindle seeds lectin to glycoconjugates of mammalian tissues showed that both lectins bind to the same histological structures. However, bark lectin has an advantage over seed lectin in a simpler purification method (lack of seed degreasing procedure) and in the possibility of harvesting raw materials for a whole year. The dynamics of changes in the hemagglutinating activity of lectins in the organs in the European spindle during the annual vegetative cycle of the plant was also studied, which made it possible to choose the optimal time for harvesting raw materials for the purification of this lectin.