Biopolym. Cell. 2020; 36(1):3-12.
Structure and Function of Biopolymers
The lectin of the bark of European spindle (Euonymus europaeus L.): purification, physico-chemical characteristics and application in histochemical studies
1Vrubel O. R., 1Sogomonian E. A., 1, 2Antonyuk V. O.
  1. Danylo Halytsky Lviv National Medical University
    69, Pekarska Str., Lviv, Ukraine, 79010
  2. Institute of Cell Biology, NAS of Ukraine
    14/16, Drahomanov Str., Lviv, Ukraine, 79005

Abstract

Aim. 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 har-vesting 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.
Keywords: European spindle, Euonymus europaeus L., lectins, carbohydrate specificity, purification, properties

References

[1] Fouquaert E, Peumans WJ, Smith DF, Proost P, Savvides SN, Van Damme EJ. The "old" Euonymus europaeus agglutinin represents a novel family of ubiquitous plant proteins. Plant Physiol. 2008;147(3):1316-24.
[2] Petryniak J, Pereira ME, Kabat EA. The lectin of Euonymus europeus: purification, characterization, and an immunochemical study of its combining site. Arch Biochem Biophys. 1977;178(1):118-34.
[3] Fouquaert E, Van Damme EJ. Promiscuity of the euonymus carbohydrate-binding domain. Biomolecules. 2012;2(4):415-34.
[4] Agostino M, Velkov T, Dingjan T, Williams SJ, Yuriev E, Ramsland PA. The carbohydrate-binding promiscuity of Euonymus europaeus lectin is predicted to involve a single binding site. Glycobiology. 2015;25(1):101-14.
[5] Shangguan W, Fan C, Chen X, Lu R, Liu Y, Li Y, Shang Y, Yin D, Zhang S, Huang Q, Li X, Meng W, Xu H, Zhou Z, Hu J, Mo X. Endothelium originated from colorectal cancer stem cells constitute cancer blood vessels. Cancer Sci. 2017;108(7):1357-1367.
[6] Toyoda M, Yamazaki-Inoue M, Itakura Y, Kuno A, Ogawa T, Yamada M, Akutsu H, Takahashi Y, Kanzaki S, Narimatsu H, Hirabayashi J, Umezawa A. Lectin microarray analysis of pluripotent and multipotent stem cells. Genes Cells. 2011;16(1):1-11.
[7] Wang S, Fan Z, Zhou B, Wang Y, Du P, Tan W, Lammi MJ, Guo X. Roles of Glycoproteins in the Diagnosis and Differential Diagnosis of Chronic and Latent Keshan Disease. Molecules. 2017;22(5). pii: E746.
[8] Antonyuk VA. Lectins and their raw materials sources. Lviv: “Quart”, 2005; 554 P.
[9] Antonyuk VA A.S. USSR No. 1554961, A method of obtaining an affinity sorbent for purification of lectins IPC B 01 J 20/22, C 12 N 11/00, publ. 04.07.90 BI No. 13, 1990.
[10] Maurer G. Disk-Electrophoresis. Moscow. “Mir”, 1971; 55–102.
[11] Lutsik AD, Detjuk ES, Lutsik MD. Lectins in Histochemistry. Lviv: “Lviv University edition”. 1989; 144 p.
[12] Van den Bergh KP, Rougé P, Proost P, Coosemans J, Krouglova T, Engelborghs Y, Peumans WJ, Van Damme EJ. Synergistic antifungal activity of two chitin-binding proteins from spindle tree (Euonymus europaeus L.). Planta. 2004;219(2):221-32.
[13] Wheatley RM, Poole PS. Mechanisms of bacterial attachment to roots. FEMS Microbiol Rev. 2018;42(4):448-461.
[14] Lannoo N, Van Damme EJ. Lectin domains at the frontiers of plant defense. Front Plant Sci. 2014;5:397.