Abstract
Lectins and its A- and B-chains from Korean mistletoe (Viscum album var.coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to ε-amino groups of lysine residues of spacer (BSA) after reduction by NaCNBH3. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A-chain and a 35kD of B-chain. Amino acid analysis andN-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturational red shift which also correspond to the conformations and activity of lectin.
Similar content being viewed by others
References Cited
Büssing, A., Suzar, K., Bergmann, J., Pf ller, U., Schietzel, M. and Schweizer, K., Induction of apoptosis in human lymphocytes treated withViscum album L. is mediated by the mistletoe lectins.Cancer Lett., 99, 59–72 (1996).
Büssing, A., Regnary, A. and Schweizer, K. Effects ofViscum album L. on cyclophosphamide-treated peripheral blood mononuclear cellsin vitro: sister chromatid exchanges and activation/proliferation maker expression.Cancer Lett., 94, 199–205 (1995).
Dietrich, J. B., Ribereau-Gayon, G., Jung, M. L., Franz, H., Beck, J.P. and Anton, R. Identity of theN-terminal sequences of the three A chains of misletoe (Viscum album L.) lectin: homology with ricin-like plant toxins and single chain ribosome-inhibiting proteins.Anticancer Drugs, 3, 507–511 (1992).
Evans, M. R. and Preece, A. W.,Viscum album-a possible treatment for cancer?Bristol Med. Chir. J. 88, 17–20 (1973).
Franz, H., Ziska P. and Kindt A., Isolation and properties of three lectins from mistletoe (Viscum album L.).Biochem. J., 195, 481–484 (1981).
Franz, H., Mistletoe lectins and their A and B chains.Oncology, 43 (suppl. 1), 23–34 (1986).
Hajto, T., Hostanska, K. and Gabius, H. J., Modulatory potency of the β-galactoside-specific lectin from mistletoe extract (Iscador) on the host defense systemin vivo in rabbits and patients.Cancer Res., 49, 4803–4808 (1989).
Hajto, T., Hostanska, K., Frey, K., Rordorf, C. and Gabius, H.J. Increased secretion of tumor necrosis fator α, interleukin 6 by human mononuclear cells exposed to β-galactoside-specific lectin from clinically applied mistletoe extract.Cancer Res., 50, 3322–3326 (1990).
Jung, M.-L., Baudino, S. and Beck J-P., Characterization of cytotoxic proteins from mistletoe (Viscum album L.).Cancer Lett., 51, 103–108 (1990).
Khwaja T. A., Dias C. B., Papoian, T. and Pentecost, S., Studies on cytotoxic and immunological effects ofViscum album (mistletoe).Proc. Am. Assoc. Cancer Res., 22, 253 (1981).
Khwaja, T. A., Dias C. B. and Pentecost S., Recent studies on the anticancer activities of mistletoe (Viscum album) and its alkaloids.Oncology, 43, suppl. 1, 42–50 (1986).
Manjikian, S., Pentecost, S. and Khwaja T.A., Isolation of cytotoxic proteins fromViscum album coloratum. Proc. Am. Ass. Cancer Res., 27 (1986).
Metzner, G., Franz, H., Kindt, A., Fahlbush, B. and Süss, J. The in vitro activity of lectin 1 from mistletoe (ML I) and its isolated A- and B-chains on functions of macrophages and polymorphonuclear cells.Immunology, 169, 461–471 (1985).
Müller, E. A. and Anderer, F. A., AViscum album oligisaccharide activating human natural cytotoxicity is an interferon gamma inducer.Cancer Immunol. Immunother., 32, 221–227 (1990).
Park, W. B. and Kim, H. S., Isolation and characterization of lectin fromViscum coloratum.Yakhak Hoeji, 38, 418–424 (1994a).
Park, W. B. and Kim, H. S., Changes of lectin fromViscum coloratum by fermentation withLactobacillus plantarum- Isolation and purification.Yakhak Hoeji, 38, 687–695 (1994b).
Park, W. B., Kim, H. S., Na, H. B. and Ham S. S., Changes of lectin fromViscum coloratum by fermentation withLactobacillus plantarum-Effects of pH and temperature, sugar specificity and lymphocyte stimulating activity.Yakhak Hoeji, 39, 24–30 (1995).
Park, W. B., Han S. K. and Han, K. H., Isolation and characterization of lectins from stem and leaves of Korean mistletoe (Viscum album coloratum) by affinity chromatography.Arch. Pham. Res. 20, 306–312 (1997).
Pfüller, U., Kopp, J., Körner I.-J., Zwanzig, M., Gockeritz, W. and Franz, H., Immunotoxins with mistletoe lectin 1 A-chain directed against interleukin-2 receptor of human lymphocytes: comparison of efficiency and specificity. In Driessche, E., Franz, H., Beeckmans, S., Pf ller, U., Kallikorm, A. and Bög-Hansen T.C. (Eds.).Lectins: Biology, Biochemistry, Clinical Biochemistry, published by Textop. Hellerup, Denmark, Vol 8, pp. 34–40, 1993.
Ribéreau-Gayon, G., Jung, M.-L., Scala, D. D. and Beck J.-P., Comparison of the effects of fermented and unfermented mistletoe preparations of cultured tumor cell.Oncology, 43 (suppl. 1), 35–41 (1986).
Ribéreau-Gayon, G., Jung, M. L., Dietrich, J. B. and Beck, J. P., Lectins and viscotoxins from mistletoe (Viscum album L.) extracts: development of a bioassay of lectins. In Driessche, E., Franz, H., Beeckmans, S., Pfüller, U., Kallikorm, A. and Bög-Hansen T.C. (Eds.),Lectins: Biology, Biochemistry, Clinical Biochemistry, published by Textop. Hellerup, Denmark, Vol 8, pp. 21–28, 1993.
Stirpe, F., Legg, R. F., Onyon, L. J., Ziska, P. and Franz, H., Inhibition of protein synthesis by a toxic lectin fromViscum album L. (Mistletoe).Biochem. J., 190, 843–845 (1980).
Schultze, J. L., Stettin, A. and Berg, P. A., Demonstration of specifically sensitized lymphocytes in patients treated with an aqueous mistletoe extract (Viscum album L.)Klin. Wonchenschr., 69, 397–403 (1991).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Park, W.B., Ju, Y.J. & Han, S.K. Isolation and characterization of β-galactoside specific lectin from Korean mistletoe (Viscum album var.coloratum) with lactose-BSA-Sepharose 4B and changes of lectin conformation. Arch. Pharm. Res. 21, 429–435 (1998). https://doi.org/10.1007/BF02974638
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02974638