Abstract
Extracts of bean (Phaseolus vulgaris L. cv. Greensleeves) cotyledons contained two abundant proteins: vicilin and phytohemagglutinin. Vicilin, a 6.9 S protein fraction at neutral pH, associated to an 18.0 S form at pH 4.5 and had 3 non-identical subunits with molecular weights (MW) of 52,000, 49,000 and 46,000. Phytohemagglutinin, a 6.4 S protein fraction, had 2 non-identical subunits with MW of 34,000 and 36,000. Phytohemagglutinin could be separated by isoelectrofocusing into a mitogenic and non-erythroagglutinating protein with a single subunit of MW=34,000, and a mitogenic and erythroagglutinating protein fraction which contained both subunits. Vicilin is apparently identical with the so called glycoprotein II (A. Pusztai and W.B. Watt, Biochim. Biophys. Acta 365, 57–71, 1970) and with globulin G1 (R.C. McLeester, T.C. Hall, S.M. Sun, F.A. Bliss, Phytochem. 2, 85; 1973), while phytohemagglutinin is identical with globulin G2 (McLeester et al., 1973). Since vicilin and phytohemagglutinin are internationally used names there is no need to introduce new names to describe P. vulgaris reserve proteins. Both proteins are catabolized in the course of seedling growth and are located in the protein bodies, indicating that they are reserve proteins. Vicilin isolated in its 18.0 S form from the cotyledons of young seedlings contains substantial quantities of smaller polypeptides, in addition the 3 original ones. We suggest that the presence of these small polypeptides represents partial breakdown of the vicilin prior to its complete catabolism.
Similar content being viewed by others
References
Allan, D., Crumpton, M.J.: Fractionation of the phytohemagglutinin of Phaseolus vulgaris by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. Biochem. Biophys. Res. Commun. 44, 1143–1148 (1971)
Barker, R.D.J., Derbyshire, E., Yarwood, A., Boulter, D.: Purification and characterization of the major storage proteins of Phaseolus vulgaris seeds, and their intracellular and cotyledonary distribution. Phytochemistry 15, 751–757 (1976)
Basha, S.M.M., Beevers, L.: The development of proteolytic activity and protein degradation during the germination of Pisum sativum L. Planta 124, 77–87 (1975)
Baumgartner, B., Chrispeels, M.J.: Purification and characterization of vicilin peptidohydrolase, the major endopeptidase in the cytoledons of mung bean seedlings. Eur. J. Biochem. 77, 223–233 (1977)
Catsimpoolas, N., Campbell, T.G., Meyer, E.W.: Immunochemical study of changes in reserve protein in germinating soybean seeds. Plant Physiol. 43, 799–805 (1968)
Danielsson, C.E.: Seed globulins of the Gramineae and Leguminosae. Biochem. J. 44, 387–400 (1949)
Daussant, J., Neucère, N.J., Conkerton, E.J.: Immunochemical studies of Arachis hypogaea proteins with particular reference to the reserve proteins. II. Protein modification during germination. Plant Physiol. 44, 480–484 (1969)
Derbyshire, E., Boulter, D.: Isolation of legumin-like protein from Phaseolus aureus and Phaseolus vulgaris. Phytochemistry 15 411–414 (1976)
Derbyshire, E., Wright, D.J., Boulter, D.: Legumin and vicilin, storage proteins of legume seeds. Phytochemistry 15, 3–24 (1976)
Ericson, M.E., Chrispeels, M.J.: Isolation and characterization of glucosamine-containing storage glycoproteins from the cotyledons of Phaseolus aureus. Plant Physiol. 52, 98–104 (1973)
Ericson, M.E., Chrispeels, M.J.: The carbohydrate moiety of mung bean vicilin. Aust. J. Plant Physiol. 3, 763–769 (1976)
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970)
Leavitt, R.D., Felsted, R.D., Bachur, N.R.: Biological and biochemical properties of Phaseolus vulgaris isolectins. J. Biol. Chem. 252, 2961–2966 (1977)
Liener, I.E.: Phytohemagglutinins (phytolectins). Ann. Rev. Plant Physiol. 27, 291–319 (1976)
Lowry, O.M., Rosebrough, M.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Manen, J.F., Miège, M.N.: Purification et caractèrisation des lectines isolées dans les albumines et les globulines de Phaseolus vulgaris Physiol. Vég. 15, 163–173 (1977)
McLeester, R.C., Hall, T.C., Sun, S.M., Bliss, F.A.: Comparison of globlin proteins from Phaseolus vulgaris with those from Vicia faba. Phytochemistry 2, 85–93 (1973)
Miller, J.B., Hsu, R., Heinrikson, R., Yachnin, S.: Extensive homology between the subunits of the phytohemagglutinin mitogenic proteins derived from Phaseolus vulgaris. Proc. Natl. Acad. Sci. USA 72, 1388–1391 (1975)
Oh, Y.H., Conard, R.A.: Further studies on mitogenic components of Phaseolus vulgaris phytohemagglutinin: subunit structure. Arch. Biochem. Biophys. 152, 631–637 (1972)
Pusztai, A., Croy, R.R.D., Grant, G., Watt, W.B.: Compartmentalization in the cotyledonary cells of Phaseolus vulgaris L. seeds: a differential sedimentation study. New Phytol. 79, 61–71 (1977)
Pusztai, A., Watt, W.B.: Glycoprotein II. The isolation and characterization of a major antigenic and non-haemagglutinating glycoprotein from Phaseoulus vulgaris. Biochim. Biophys. Acta 207, 413–431 (1970)
Pusztai, A., Watt, W.B.: Isolectins of Phaseolus vulgaris. A comprehensive study of fractionation. Biochim. Biophys. Acta 365, 57–71 (1974)
Racusen, D., Foote, M.: The major glycoprotein in germinating bean seeds. Can. J. Bot. 49, 2107–2111 (1971)
Sun, S.M., McLeester, R.C., Bliss, F.A., Hall, T.C.: Reversible and irreversible dissociation of globulins from Phaseolus vulgaris seeds. J. Biol. Chem. 249, 2218–2221 (1974)
Thanh, V.H., Okubu, K., Shibasaki, K.: Isolation and characterization of the multiple 7 S globulins of soybean proteins. Plant Physiol. 56, 19–22 (1975)
Thomson, J.A., Schroeder, H.H., Dudman, W.F.: Cotyledonary storage proteins in Pisum sativum. I. Molecular heterogeneity. Aust. J. Plant Physiol. (in press)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bollini, R., Chrispeels, M.J. Characterization and subcellular localization of vicilin and phytohemagglutinin, the two major reserve proteins of Phaseolus vulgaris L.. Planta 142, 291–298 (1978). https://doi.org/10.1007/BF00385080
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00385080