Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Phaseolus vulgaris phytohemagglutinin contains high-mannose and modified oligosaccharide chains


Phytohemagglutinin, the major lectin in the seeds of the common bean Phaseolus vulgaris L., was isolated by affinity chromatography from cotyledons of nearly mature seeds and from developing cotyledons labeled with [3H]glucosamine, [3H]mannose or [3H]fucose. The protein was subjected to exhaustive proteolysis and the carbohydrate composition of the resulting glycopeptides examined. Two classes of oligosaccharide side-chains were found. The sidechains of the first class are of the high-mannose type, containing two residues of N-acetylglucosamine and 8 or 9 mannose residues. The sidechains of the second class are of the modified type containing N-acetylglucosamine, mannose, fucose, xylose in molar ratios of 2:3.8:0.6:0.5. Two-dimensional gel electrophoresis shows that phytohemagglutinin can be fractionated into seven different glycosylated polypeptides, and that each one contains at least one modified oligosaccharide chain. The results indicate that most glycosylated polypeptides probably contain one chain of each class. The carbohydrate composition of the two types of chains is similar to that found in other plant glycoproteins, but this is the first report of a plant glycoprotein with both highmannose and modified oligosaccharides on the same polypeptide chain.

This is a preview of subscription content, log in to check access.


endo H:

endo-β-N-acetylglucosaminidase H










  1. Allen, L.W., Svenson, R.H., Yachnin, S. (1969) Purification of mitogenic proteins derived from Phaseolus vulgaris: isolation of potent and weak phytohemagglutinins possessing mitogenic activity. Proc. Natl. Acad. Sci. USA 63, 334–341

  2. Badenoch-Jones, J., Spencer, D., Higgins, T.J.V., Millerd, A. (1981) The role of glycosylation in storage protein synthesis in developing pea seeds. Planta 153, 201–209

  3. Bollini, R., Chrispeels, M.J. (1978) Characterization and subcellular localization of vicilin and phytohemagglutinin, the two major reserve proteins of Phaseolus vulgaris L. Planta 142, 291–298

  4. Bollini, R., Van der Wilden, W., Chrispeels, M.J. (1982) A precursor of the reserve-protein phaseolin is transiently associated with the endoplasmic reticulum of developing Phaseolus vulgaris cotyledons. Physiol. Plant 55, 82–92

  5. Chrispeels, M.J. (1983a) Incorporation of fucose into the carbohydrate moiety of phytohemagglutinin in developing cotyledons of Phaseolus vulgaris. Planta 157, 454–461

  6. Chrispeels, J.J. (1983b) The Golgi apparatus mediates the transport of phytohemagglutinin to the protein bodies in bean cotyledons. Planta 158, 140–151

  7. Chrispeels, M.J., Bollini, R. (1982) Characteristics of membrane-bound lectin in developing Phaseolus vulgaris cotyledons. Plant Physiol. 70, 1425–1428

  8. Clarke, J., Shannon, L.M. (1976) The isolation and characterization of the glycopeptides from horseradish peroxidase isoenzyme C. Biochim. Biophys. Acta 427, 428–442

  9. Davies, H.M., Delmer, D.P. (1979) Seed reserve-protein glyco-sylation in an in vitro preparation from developing cotyledons of Phaseolus vulgaris. Planta 146, 513–520

  10. Davies, H.M., Delmer, D.P. (1981) Two kinds of protein glycosylation in a cell-free preparation from developing cotyledons of Phaseolus vulgaris. Plant Physiol. 68, 284–291

  11. Ericson, M.C., Delmer, D. (1977) Glycoprotein synthesis in plants. I. Role of lipid intermediates. Plant Physiol. 59, 341–347

  12. Felsted, R.L., Laevitt, R.D., Bachur N.R. (1975) Purification of the phytohemagglutinin family of proteins from red kidney beans (Phaseolus vulgaris) by affinity chromatography. Biochim. Biophys. Acta 405, 72–81

  13. Foresee, W.T., Elbein, A.D. (1975) Glycoprotein biosynthesis in plants. Demonstration of lipid-linked oligosaccharides of mannose and N-acetylglucosamine. J. Biol. Chem. 250, 9283–9293

  14. Hori, H., James, D.W., Elbein, A. (1982) Isolation and characterization of the Glc3 Man9 GlcNAc2 from lipid-linked oligosaccharides of plants. Arch. Biochem. Biophys. 215, 12–21

  15. Hubbard, S.C., Ivatt, R.J. (1981) Synthesis and processing of asparagine-linked oligosaccharides. Annu. Rev. Biochem. 50, 555–583

  16. Iglesias, J.L., Lis, H., Sharon, N. (1982) Purification and properties of a D-galactose/N-acetyl-D-galactosamine-specific lectin from Erythrina crystagalli. Eur. J. Biochem. 123, 247–252

  17. Ishihara, H., Takahashi, N., Oguri, S., Tejima, S. (1979) Complete structure of the carbohydrate moiety of stem bromelain. J. Biol. Chem. 254, 10715–10719

  18. Jaffé, W.G., Levy, A., Gonzales, D.I. (1974) Isolation and partial characterization of bean phytohemagglutinin. Phytochemistry 13, 2685–2693

  19. Kornfeld, S. (1982) Oligosaccharide processing during glycoprotein biosynthesis. In: The glycoconjugates, vol. 3, pp. 3–23, Horowitz, M., ed. Academic Press, New York

  20. Kurokawa, T., Tsuda, M., Sugino, Y. (1976) Purification and characterization of a lectin from Wistaria floribunda seeds. J. Biol. Chem. 251, 5686–5693

  21. Leavitt, R.D., Felsted, R.L., Bachur, N.R. (1977) Biological and biochemical properties of Phaseolus vulgaris isolectins. J. Biol. Chem. 252, 2961–2966

  22. Ledeen, R.W., Yu, R.K. (1982) Gangliosides: structure, isolation, and analysis. Methods Enzymol. 83, 139–180

  23. Lehle, L. (1981) Plant cells synthesize glucose-containing lipid-linked oligosaccharides similar to those found in animals and yeast. FEBS Lett. 123, 63–66

  24. Lehnhardt, W.F., Winzler, R.J. (1968) Determination of neutral sugars in glycoproteins by gas-liquid chromatography. J. Chromatogr. 34, 471–479

  25. Li, E., Kornfeld, S. (1979) Structural studies of the major high-mannose oligosaccharide units from Chinese hamster ovary cell glycoproteins. J. Biol. Chem. 254, 1600–1605

  26. Mialonier, G., Privat, J.-P., Monsigny, M., Kahlem, G., Durand, R. (1973) Isolement, propriétés physico-chimiques et localisation in vivo d'une phytohémagglutinine (lectine) de Phaseolus vulgaris L. (var. rouge). Physiol. Veg. 11, 519–537

  27. Misaki, A., Goldstein, I.J. (1977) Glycosyl moiety of the lima bean lectin. J. Biol. Chem. 252, 6995–6999

  28. O'Farrel, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021

  29. Ohtani, K., Misaki, A. (1980) The structure of the glycan moiety of Tora bean (Phaseolus vulgaris). Carbohydr. Res. 87, 275–285

  30. Räsänen, V., Weber, T.H., Gräsbeck, R. (1973) Crystalline kidney-bean leucoagglutinin. Eur. J. Biochem. 38, 193–200

  31. Spencer, D., Higgins, T.J.V., Button, S.C., Davey, R.A. (1980) Pulse-labeling studies on protein synthesis in developing pea seeds and evidence of a precursor form of legumin small subunit. Plant Physiol. 66, 510–515

  32. Staneloni, R.J., Tolmasky, M.E., Petriella, C., Ugalde, R.A., Leloir, L.F. (1980) Presence in a plant of a compound similar to the dolichyldiphosphate oligosaccharide of animal tissue. Biochem. J. 191, 257–260

  33. Tarentino, A.L., Maley, F. (1974) Purification and properties of an endo-β-N-acetylglucosaminidase from Streptomices griseus. J. Biol. Chem. 249, 811–817

  34. Van der Wilden, W., Herman, E., Chrispeels, M.J. (1980) Protein bodies of mung bean cytoledons as autophagic organelles. Proc. Natl. Acad. Sci. USA 77, 428–432

  35. Warner, T.G., O'Brien, J.S. (1982) Structure analysis of the major oligosaccharides accumulating in canine GM1 gangliosidosis liver. J. Biol. Chem. 257, 224–232

Download references

Author information

Correspondence to Maarten J. Chrispeels.

Additional information

This work was done while A.V. was on leave from the Istituto Biosintesi Vegetali, C.N.R., via Bassini 15, I-20133 Milano, Italy

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Vitale, A., Warner, T.G. & Chrispeels, M.J. Phaseolus vulgaris phytohemagglutinin contains high-mannose and modified oligosaccharide chains. Planta 160, 256–263 (1984).

Download citation

Key words

  • Asparagine linked oligosaccharides
  • Cotyledon
  • Glycoprotein
  • Lectin
  • Phaseolus (lectin structure)
  • Phytohemagglutinin (structure)