Abstract
A crude organelle preparation from germinating castor bean endosperm catalysed the incorporation of mannose from GDP[14C]mannose into acid-labile mannolipids. Solubility and chromatographic properties have identified the most rapidly synthesized products as mannosyl-phosphoryl-polyisoprenol, while the more polar lipid formed was shown to contain oligosaccharide. Little radioactivity from GDP[14C]mannose accumulated in insoluble product in the cell-free system, but supplying GDP[14C]mannose to intact endosperm tissue has shown that the major incorporation product in vivo is glycoprotein. This product was readily solubilized by either pronase or sodium dodecyl sulphate treatment suggesting it was membrane bound glycoprotein. Incorporation of mannose into mannosyl-phosphoryl-polyisoprenol during the cell-free assay was stimulated by the addition of dolichol monophosphate. This enzymic activity was optimal at pH 7.5 and in the presence of 10 mM Mg2+. The Km for GDP-mannose was estimated to be 5×10-7 M. Cellular mannosyl transferase activity changed markedly during early post-germinative growth; from being absent in the dry seed, enzyme activity increased to peak between the second and third days of growth and subsequently declined.
Similar content being viewed by others
Abbreviations
- TCA:
-
trichloroacetic acid
- SDS:
-
sodium dodecyl sulphate
References
Alam, S.S., Hemming, F.W.: Polyprenol phosphates and mannosyl transferase in Phaseolus aureus. Phytochemistry 12, 1641–1649 (1973)
Babczinski, P., Tanner, W.: Involvement of dolichol monophosphate in the formation of specific mannosyl-linkages in yeast glycoproteins. Biochem. Biophys. Res. Commun. 54, 1119–1124 (1973)
Baynes, J.W., Hsu, A.F., Heath, E.C.: The role of mannosyl-phosphoryl-dihydropolyisoprenol in the synthesis of mammalian glycoproteins. J. Biol. Chem. 248, 5693–5704 (1973)
Beevers, H.: Glyoxysomes from castor bean endosperm and their relation to glyconeogenesis. Ann. N.Y. Acad. Sci. 168, 313–324 (1969)
Beevers, L., Mense, R.M.: Glycoprotein biosynthesis in cotyledons of Pisum savieum L. Involvement of lipid-linked intermediates. Plant Physiol. 60, 703–708 (1977)
Behrens, N.H., Parodi, A.J., Leloir, L.F.: Glucose transfer from dolichol monophosphate glucose: the product formed with endogenous microsomal acceptor. Proc. Nat. Acad. Sci. USA 68, 2857–2860 (1971)
Behrens, N.H., Carminatti, H., Staneloni, R.J., Leloir, L.F., Cantarella, A.I.: Formation of lipid-bound oligosaccharides containing mannose. Their role in glycoprotein biosynthesis. Proc. Nat. Acad. Sci. USA 70, 3390–3394 (1973)
Bowden, L., Lord, J.M.: The cellular origin of glyoxysomal proteins in germinating castor bean endosperm. Biochem. J. 154, 501–506 (1976)
Bray, G.A.: A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Anal. Biochem. 1, 270–285 (1960)
Erieson, M.C., Chrispeels, M.J.: Isolation and characterization of glucosamine containing glycoproteins from the cotyledons of Phaseolus aureus. Plant Physiol. 52, 98–104 (1973)
Ericson, M.C., Delmer, D.P.: Glycoprotein synthesis in plants. I. Role of lipid intermediates. Plant Physiol. 59, 341–347 (1977)
Ericson, M.C., Delmer, D.P.: Glycoprotein synthesis in Plants. III. Interaction between UDP-N-acetylglucosamine and GDP-mannose as substrates. Plant Physiol. 61, 819–823 (1978)
Forsee, W.T., Elbein, A.D.: Glycoprotein biosynthesis in plants. Demonstration of lipid-linked oligosaccharides of mannose and N-acetyl glucosamine. J. Biol. Chem. 250, 9283–9293 (1975)
Forsee, W.T., Valkovitz, G., Elbein, A.D.: Glycoprotein biosynthesis in plants. Formation of lipid-linked oligosaccharides of mannose and N-acetylglucosamine by mung bean seedling. Arch. Biochem. Biophys. 174, 469–479 (1976)
Green, J.R., Northcote, D.H.: The structure and function of glycoproteins synthesized during slime-polysaccharide production by membranes of the root-cap cells of maize (Zea mays). Biochem. J. 170, 599–608 (1978)
Hsu, A.F., Baynes, J.W., Heath, E.C.: The role of a dolichol-oligosaccharide as an intermediate in glycoprotein biosynthesis. Proc. Nat. Acad. Sci. USA 71, 2391–2395 (1974)
Kauss, H.: A plant mannosyl-lipid acting in reversible transfer of mannose. FEBS Lett. 5, 81–84 (1969)
Kiely, M.L., McKnight, S., Schimke, R.T.: Studies on the attachment of carbohydrate to ovalbumin nascent chains in hen oviduct. J. Biol. Chem. 251, 5490–5495 (1976)
Lamport, D.T.A.: Cell wall metabolism. Ann. Rev. Plant Physiol. 31, 235–270 (1970)
Lehle, L., Tanner, W.: Formation of lipid-bound oligosaccharides in yeast. Biochim. Biophys. Acta 399, 364–374 (1975)
Lehle, L., Fartaczek, F., Tanner, W., Kass, H.: Formation of polyprenol-linked mono- and oligosaccharides in Phaseolus aureus. Arch. Biochem. Biophys. 175, 419–426 (1976)
Lennarz, W.J.: Lipid-linked sugars in glycoprotein synthesis. Science 188, 986–991 (1975)
Lingappa, V.R., Lingappa, J.R., Prasad, R., Ebner, K.E., Blobel, G.: Coupled cell-free synthesis, segregation and core glycosylation of a secretory protein. Proc. Nat. Acad. Sci. USA 75, 2338–2342 (1978)
Lucas, J.J., Waechter, C.J.: Polyisoprenoid glycolipids involved in glycoprotein biosynthesis. Mol. Cell. Biochem. 11, 67–78 (1978)
Lucas, J.J., Waechter, G.J., Lennarz, W.J.: The participation of lipid-linked oligosaccharide in synthesis of membrane glycoproteins. J. Biol. Chem. 250, 1992–2002 (1975)
Mellor, R.B., Bowden, L., Lord, J.M.: Glycoproteins of the glyoxysomal matrix. FEBS Lett. 90, 275–278 (1978)
Oliver, G.J.A., Hemming, F.W.: The transfer of mannose to dolichol diphosphate oligosaccharides in pig liver endoplasmic reticulum. Biochem. J. 152, 191–199 (1975)
Riezman, H., Mersey, B.G., Lamb, J.E., Becker, W.M.: Detection and distribution of glyoxysomal glycoproteins. Plant Physiol. 61, 105 (1978)
Trevelyan, W.E., Proctor, D.P., Harrison, J.S.: Detection of sugars on paper chromatograms. Nature (London) 166, 444–445 (1950)
Waechter, C.J., Lennarz, W.J.: The role of polyprenol-linked sugars in glycoprotein synthesis. Ann. Rev. Biochem. 45, 95–112 (1976)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mellor, R.B., Lord, J.M. Formation of lipid-linked mono- and oligosaccharides from GDP-mannose by castor bean endosperm homogenates. Planta 146, 91–99 (1979). https://doi.org/10.1007/BF00381260
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00381260