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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Purification and Partial Characterization of a Lectin from the Fresh Leaves of Kalanchoe crenata (Andr.) Haw

  • Published : 2004.03.31
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Abstract

A haemagglutinating protein from the saline extracts of Kalanchoe crenata leaves, which agglutinate all human blood types, was purified to homogeneity by ion-exchange chromatography on a DEAE-Cellulose column followed by gel filtration on a Sephadex G-100 column. The purified protein showed one band, both in non-denaturing PAGE and SDS-PAGE. The $M_r$ that was determined by SDS-PAGE was 44,000 Da and that estimated from gel filtration was 47,000. Treatment of the haemagglutinating protein with 5 mM EDTA diminished the haemagglutinating activity to 50% of the original level. The addition of divalent cations, 10 mM $Mg^{2+}$, 10 mM $Mn^{2+}$, or 10 mM $Ba^{2+}$, totally restored and enhanced the activity. The protein showed maximum activity over the 3-7 pH range and was heat-resistant. It was also a glycoprotein containing about 1.5% carbohydrate.

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References

  1. Adeyemi, A., Togun, R. A., Ogunbiyi, O. A. and Aboderin A. (1994) Observations on the Haemagglutinins from the seeds of Dioclea reflexa (Hook). Nigerian J. Biochem. 9, 26-36.
  2. Adolph, L. and Lorenz, R. (1982) Enzyme Diagnosis in Diseases of the Heart, Liver and Pancreas, John Wiley and Sons Inc., New York, USA.
  3. Anuradha, P. and Bhide, S. V. (1999). An isolectin complex from Tricosanthes anguina seeds. Phytochem. 52, 751-758. 1. https://doi.org/10.1016/S0031-9422(99)00288-5
  4. Bing, D. H., Weyand, J. G. M. and Stavitsky, A. B. (1967) Heamagglutination with aldehyde fixed erythrocytes for assay of antigens and antibodies. Proc. Soc. Exp. Biol. Med. 124, 1166-1170. https://doi.org/10.3181/00379727-124-31953
  5. Cavada, B. S., Santos, C. F., Grangeiro, B. T., Ramos, R. L. and Calvete, J. J. (1998) Purification and characterization of a lectin from seeds of Vatainea macrocarpa. Phytochem. 49, 675-680 https://doi.org/10.1016/S0031-9422(98)00144-7
  6. Dalziel, J. M. (1955) The Useful Plants of West Tropical Africa, Millibank, London, UK.
  7. Dong, T. X., Ng, T. B., Wong, R. N. S., Yeung, H. W. and Xu, G. J. (1993) Investigation of haemagglutinating activity in seeds of various Tricosanthes Species. Internat. J. Biochem. 25, 411-414. https://doi.org/10.1016/0020-711X(93)90632-O
  8. Goldstein, I. J. and Poretz, R. D. (1986) Isolation, physicochemical characterization and carbohydrate-binding specificity of lectins; in The Lectins: Properties, Functions and Applications in Biology and Medicine, Liener, I. E., Sharon, N. and Goldstein, I. J. (eds.), Academic Press Inc., Orlando, USA.
  9. Gornall, A. G., Bardwill, C. J. and David, M. M. (1949) Determination of serum proteins by Biuret reaction. J. Biol. Chem. 177, 751-766.
  10. Hayes, C. E. and Goldstein, I. J. (1974) An $\alpha$-D-galactosylbinding lectin from Bandeiraea simplicifolia seeds-isolation by affinity chromatography and characterization. J. Biol. Chem. 249, 1904-1914.
  11. Jermyn, M. A. (1975) Increasing the sensitivity of the anthrone method for carbohydrate analysis. Biochemistry 68, 322-335.
  12. Knight, C. S. (1967) Improved techniques with advanced ionexchange cellulose. Adv. Chromat. 4, 61-63.
  13. Kuku, A., Stoppini, M., Cobianchi, A., Minetti, G., Balduini, C. and Aboderin, A. (2000) The complete primary structure of a mannose/glucose specific lectin from the seeds of Dioclea reflexa (Hook, F.) Nig. J. Biochem. Mol. Biol. 15, 115-119.
  14. Lis, H. and Sharon, N. (1989) Lectins in higher plants; in Biochemistry of Plants, Vol. 6, pp. 371-447. Academic Press Inc, New York, USA.
  15. Lotan, R., Skutelsky, E., Danon, D and Sharon, N. (1975) The purification, composition and specificity of the antiT lectin from peanut (Arachea hypogea). J. Biol. Chem. 250, 8518-8523.
  16. Moreira, A. R., Castelo-Branco, C. C, Monteiro, A. C. O., Tavares, R. O. and Beltramini, L. M. (1998) Isolation and partial characterization of a lectin from Artocarpus incisa seed. Phytochem. 47, 1183-1188. https://doi.org/10.1016/S0031-9422(97)00753-X
  17. Presant, C. A. and Kornfeld, S. (1972) Characterisation of the cell surface receptor for the Agaricus bisporus heamagglutinin. J. Biol. Chem. 247, 6937-6945.
  18. Pusztai, A. (1993) Dietary lectins are metabolic signals for the gut and modulate Immune and hormone functions. Eur. J. Clin. Nutr., 47, 691-699.
  19. Sampaio, A. H., Rogers, D. S. J. and Barwell, C. J. (1998) A galactose-specific lectin from the red marine alga, Ptilota filicina. Phytochem. 48, 765-769. https://doi.org/10.1016/S0031-9422(97)00966-7
  20. Sharon, N. (1993) Lectin-carbohydrate complexes of plants and animals: An atomic view. Trends Biochem. Sci. 18, 221-226. https://doi.org/10.1016/0968-0004(93)90193-Q
  21. Shyamasri, B. and Arvind, M. K. (2002) Thermal stability of Phaseolus vulgaris leucoagglutinin: A differential scanning calorimetry. J. Biochem. Mol. Biol. 35, 472-475. https://doi.org/10.5483/BMBRep.2002.35.5.472
  22. Togun, R. A., Animashaun, T., Binutu, O. and Aboderin, A. (1994) $\beta$-Galactosidase binding lectins from the seeds of Tetracarpidium conophorum (Mull. Arg. Hutch and Dalz.) (Euphorbiaceae). Nigerian J. Biochem. 9, 45-53.
  23. Utarabhand, P. and Akkayanont, P. (1995) Purification of a lectin from Parkia javanica beans. Phytochem. 38, 281-285. https://doi.org/10.1016/0031-9422(94)00550-D
  24. Watkins, W. M., Yates, A. D. and Greenwell, P. (1981) Blood group antigens and the enzymes involved in their syntheses, past and present. Trans. Biochem. Soc. 9, 186-191
  25. Wittsuwannakul, R., Wittsuwannakul, D. and Sakulborirug, C. (1998) A lectin from the bark of the rubber tree (Hevea brasiliensis). Phytochem. 47, 183-187. https://doi.org/10.1016/S0031-9422(97)00329-4

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