Skip to main content
SpringerLink
Log in

Transport and posttranslational processing of the vacuolar enzyme α-mannosidase in jack-bean cotyledons

  • Published:
Planta Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Abstract

α-Mannosidase (EC 3.2.1.24) is a vacuolar enzyme which occurs abundantly in the cotyledons of the jack-bean (Canavalia ensiformis (L.) DC). The mature enzyme is a tetramer with two polypeptides each of relative molecular mass (Mr) 66000 and Mr 44000. The enzyme has an interesting molecular structure because in its native form, it does not bind to concanavalin A (ConA) in spite of the presence of a high-mannose glycan. α-Mannosidase is synthesized in the developing cotyledons of jack-beans at the same time as the abundant proteins canavalin and ConA. The enzyme is synthesized as a precursor which has an Mr of 110000 and is associated with the endoplasmic reticulum (ER). Antibodies against the deglycosylated subunits cross-react with the Mr-110000 precursor. Processing of the precursor to the constituent polypeptides occurs posttranslationally, probably in the protein bodies. Immunocytochemical evidence shows that α-mannosidase is present in the ER and the Golgi complex of developing cells, and accumulates in the protein bodies.

Labeling with [3H]glucosamine shows that after processing only the Mr-66000 polypeptide has glucosamine-containing glycans. The synthesis of these glycans is inhibited by tunicamycin, indicating that they are asparagine-linked oligosaccharides. Analysis of the glycans shows that there is a large glycan that is retained by ConA and a small glycan that is not retained by ConA. The large glycan is only partially sensitive to α-mannosidase because of the presence of a terminal glucose residue. Cross-reaction of the large subunit with an antiserum directed against small, complex glycans of plant glycoproteins indicates that this polypeptide probably has a xylose-containing glycan. Pulse-chase experiments carried out in the presence of tunicamycin show that the presence of glycans is not required for transport of α-mannosidase out of the ER-Golgi system.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ConA:

concanavalin A

ER:

endoplasmic reticulum

H L:

heavy, light subunit

IgG:

Immunoglobulin G

Mr :

relative molecular mass

SDS-PAGE:

sodium dodecylsulfate-polyacrylamide gel electrophoresis

References

  • Bendiak, B., Cumming, D.A. (1985) Hydrozinolysis-N-reacetylation of glycopeptides and glycoproteins. Model studies using 2-acetamido-1-N-[l-aspart-4-oyl]-2-deoxy-B-d-glucopyranosylamine. Carbohydr. Res. 144, 1–12

    Google Scholar 

  • Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132

    Google Scholar 

  • Bollini, R., Ceriotti, A., Daminati, M.G., Vitale, A. (1985) Glycosylation is not needed for the intracellular transport of phytohemagglutinin in developing Phaseolus vulgaris cotyledons and for the maintenance of its biological activities. Plant Physiol. 65, 15–22

    Google Scholar 

  • Bollini, R., Chrispeels, M.J. (1979) The rough endoplasmic reticulum is the site of reserve-protein synthesis in developing Phaseolus vulgaris cotyledons. Planta 146, 487–501

    Google Scholar 

  • Bollini, R., Vitale, A., Chrispeels, M.J. (1983) In vivo and in vitro processing of seed reserve protein in the endoplasmic reticulum: Evidence for two glycosylation steps. J. Cell Biol. 96, 999–1007

    Google Scholar 

  • Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Bicohem. 46, 83–88

    Google Scholar 

  • Borroto, K., Dure, L., III (1987) The globulin seed storage proteins of flowering plants are derived from two ancestral genes. Plant Mol. Biol. 8, 113–131

    Google Scholar 

  • Bowles, D.J., Andralojc, J., Marcus, S. (1982) Identification of an endogenous ConA-binding polypeptide as the heavy subunit of α-mannosidase. FEBS Lett. 140, 234–236

    Google Scholar 

  • Bowles, D.J., Chaplin, M.F., Marcus, S.E. (1983) Interaction of concanavalin A with native and denatured forms of jackbean α-d-mannosidase. Eur. J. Biochem. 130, 613–618

    Google Scholar 

  • Chrispeels, M.J. (1984) Biosynthesis, processing, and transport of storage proteins and lectins in legume cotyledons. Phil. Trans. R. Soc. London B304, 309–322

    Google Scholar 

  • Chrispeels, M.J. (1985) The role of the Golgi apparatus in the transport and post-translational modification of vacuolar (protein body) proteins. Oxford Surv. Plant Mol. Cell Biol. 2, 43–68

    Google Scholar 

  • Chrispeels, M.J., Higgins, T.J.V., Craig, S., Spencer, D. (1982) Role of the endoplasmic reticulum in the synthesis of reserve proteins and the kinetics of their transport to protein bodies in developing pea cotyledons. J. Cell Biol. 93, 5–14

    Google Scholar 

  • Craig, S., Goodchild, D.J., Miller, C. (1980) Structural aspects of protein accumulation in developing pea cotyledons. II. Three-dimensional reconstructions of vacuoles and protein bodies from serial sections. Aust. J. Plant Physiol. 7, 329–337

    Google Scholar 

  • Danscher, G., Norgaard, J.O.R. (1983) Light microscopic visualization of colloidal gold on resin-embedded tissue. J. Histochem. Cytochem. 31, 1394–1398

    Google Scholar 

  • Doyle, J.J., Schuler, M.A., Godette, W.D., Zeuger, V., Beachy, R.N., Slightom, J.L. (1986) The glycosylated seed storage proteins of Glycine max and Phaseolus vulgaris. Structural homologies of genes and proteins. J. Biol. Chem. 260, 6561–6564

    Google Scholar 

  • Edge, A.S.B., Faltynek, C.R., Hof, L., Reichert, L.E. Jr., Weber, P. (1981) Deglycosylation of glycoproteins by trifluoromethanesulfonic acid. Anal. Biochem. 118, 131–137

    Google Scholar 

  • Faye, L., Chrispeels, M.J. (1985) Characterization of N-linked oligosaccharides by affinoblotting with Concanavalin A-peroxidase and treatment of the blots with glycosidases. Anal. Biochem. 149, 218–244

    Google Scholar 

  • Faye, L., Chrispeels, M.J. (1987) Transport and processing of the glycosylated precursor of concanavalin A in jack-bean. Planta 170, 217–224

    Google Scholar 

  • Gatehouse, J.A., Lycett, G.W., Croy, R.R.D., Boulter, D. (1982) The post-translational proteolysis of the subunits of vicilin from pea (Pisum sativum L.). Biochem. J. 207, 629–632

    Google Scholar 

  • Graham, J.S., Pearce, G., Merryweather, J., Titani, K., Ericson, L., Ryan, C.A. (1985) Wound-induced proteinase inhibitor from tomato leaves. J. Biol. Chem. 260, 6555–6560

    Google Scholar 

  • Harris, N., Chrispeels, M.J. (1975) Histochemical and biochemical observations on storage protein metabolism and protein body autolysis in cotyledons of germinating mung beans. Plant Physiol. 56, 292–299

    Google Scholar 

  • Hase, S., Koyama, S., Daiyasu, H., Takemoto, H., Hara, S., Kobayashi, Y., Kyogoku, Y., Ikenaka, T. (1986) Structure of a sugar chain of a protease inhibitor isolated from Barbados Pride (Casesalpinia pulcherrima Sw.) seeds. J. Biochem. 100, 1–10

    Google Scholar 

  • Herman, E.M., Shannon, L.M. (1984) Accumulation and subcellular localization of α-galactosidase in developing soybean cotyledons. Plant Physiol. 77, 886–890

    Google Scholar 

  • Johnson, L.M., Bankaitis, V.A., Emr, S.D. (1987) Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease. Cell 48, 875–885

    Google Scholar 

  • Kaladas, P.M., Goldberg, R., Poretz, R.D. (1983) Rabbit anticarbohydrate antibody elicited by the lymphocyte mitogenic glycoprotein from Wistaria floribunda seeds. Molec. Immunol. 20, 727–735

    Google Scholar 

  • Kitagaki-Ogawa, H., Matsumoto, I., Seno, N., Takahashi, N., Endo, S., Arata, Y. (1986) Characterization of the carbohydrate moiety of Clerodendron trichotomum lectins. Its structure and reactivity toward plant lectins. Eur. J. Biochem. 161, 779–785

    Google Scholar 

  • Laemmli, U.K., Favre, M. (1973) Maturation of the head of bacteriophage T4. J. Mol. Biol. 80, 575–599

    Google Scholar 

  • Laurière, M., Laurière, C., Sturm, A., Chrispeels, M.J. (1987) Characteristics of the soluble and cell wall associated β-fructosidase from suspension-cultured carrot cells. (Abstr.) Plant Physiol. 83, Suppl., 91

    Google Scholar 

  • Laurière, C., Laurière, M., Sturm, A., Faye, L., Chrispeels, M.J. (1988) Characterization of β-fructosidase, an extracellular glycoprotein of carrot cells. Biochimie (in press)

  • Martinoia, E., Heck, V., Wiemken, A. (1981) Vacuoles as storage compartments for nitrate in barley leaves. Nature 289, 292–294

    Google Scholar 

  • Matile, P.H. (1975) The lytic compartment of plant cells (Cell Biology Monogr., vol. 1). Springer, New York

    Google Scholar 

  • Mellis, S.J., Baenziger, J.U. (1981) Separation of neutral oligosaccharides by high-performance liquid chromatography. Anal. Biochem. 114, 276–280

    Google Scholar 

  • Mierendorf, R.C., Cardelli, J.A., Livi, G.P., Dimond, R.L. (1983) Synthesis of related forms of the lysosomal enzyme α-mannosidase in Dictyostelium discoideum. J. Biol. Chem. 258, 5878–5884

    Google Scholar 

  • Millerd, A., Spencer, D., Dudman, W.F., Stiller, M. (1975) Growth of immature pea cotyledons in culture. Aust. J. Plant Physiol. 2, 51–59

    Google Scholar 

  • Opheim, D.J., Touster, O. (1978) Lysosomal α-d-Mannosidase of rat liver. J. Biol. Chem. 253, 1017–1023

    Google Scholar 

  • Paus, E. (1977) α-Mannosidase from Phaseolus vulgaris. Composition and structural properties. Eur. J. Biochem. 73, 155–161

    Google Scholar 

  • Schwaiger, H., Hasilik, A., von Figura, K., Wiemken, A., Tanner, W. (1982) Carbohydrate-free carboxypeptidase Y is transferred into the lysosome-like yeast vacuole. Bioch. Biophys. Res. Commun. 104, 950–956

    Google Scholar 

  • Shepherd, V., Montgomery, R. (1976) α-d-Mannosidase, preparation and properties of free and insolubilized enzyme. Biochim. Biophys. Acta 429, 884–894

    Google Scholar 

  • Slot, J.W., Gueze, H.J. (1985) A new method of preparing gold probes for multiple labeling cytochemistry. Eur. J. Cell Biol. 38, 87–93

    Google Scholar 

  • Sly, W.S., Fisher, H.D. (1982) The phosphomannosyl recognition system for intracellular and intercellular transport of lysosomal enzymes. J. Cell Biochem. 18, 67–85

    Google Scholar 

  • Spencer, D., Chandler, P.M., Higgins, T.J.V., Inglis, A.S., Rubira, M. (1983) Sequence interrelationships of the subunits of vicilin from pea seeds. Plant Mol. Biol. 2, 259–267

    Google Scholar 

  • 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

    Google Scholar 

  • Stinissen, H.M., Peumans, W.J., Chrispeels, M.J. (1984) Subcellular site of lectin synthesis in developing rice embryos. EMBO J. 3, 1979–1985

    Google Scholar 

  • Sturm, A., Chrispeels, M.J., Wieruszeski, J.M., Strecker, G., Montreuil, J. (1987) Structural analysis of the N-linked oligosaccharides from jack-bean α-mannosidase. In: Glycoconjugates, p. A107, Montreuil, J., Verbert, A., Spik, G., Fournet, B., eds. Secretariat, 9th Int. Glycoconjugate Symp., Lille, France

  • Takasaki, S., Yamashita, K., Suzuki, K., Jwanaga, S., Kobota, A. (1979) The sugar chains of cold-insoluble globulin. J. Biol. Chem. 254, 8548–8553

    Google Scholar 

  • Valls, L.A., Hunter, C.P., Rothman, J.H., Stevens, T.H. (1987) Protein sorting in yeast: the localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide. Cell 48, 887–897

    Google Scholar 

  • Van der Wilden, W., Chrispeels, M.J. (1983) Characterization of the isozymes of α-mannosidase located in the cell wall, protein bodies, and endoplasmic reticulum of Phaseolus vulgaris cotyledons. Plant Physiol. 71, 82–87

    Google Scholar 

  • Van der Wilden, W., Gilkes, N.R., Chrispeels, M.J. (1980) The endoplasmic reticulum of mung bean cotyledons: role in the accumulation of hydrolases in protein bodies during seedling growth. Plant Physiol. 66, 390–394

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Wang, B.-L., Scopsi, L., Nielsen, M.H., Larsson, L.-I. (1985) Simplified purification and testing of colloidal gold probes. Histochemistry 83, 109–115

    Google Scholar 

Download references

Author information

Author notes

  1. Loic Faye

    Present address: Faculté des Sciences de Rouen, Centre National de La Recherche Scientifique (UA 203), F-76130, Mont Saint Aignan, France

  2. John S. Greenwood

    Present address: Department of Botany, University of Guelph, N1G 2W1, Guelph, Ont., Canada

  3. Eliot M. Herman

    Present address: Plant Molecular Genetics Laboratory, U.S. Department of Agriculture, 20705, Beltsville, MD, USA

Authors and Affiliations

  1. Department of Biology, University of California San Diego, 92093-0016, La Jolla, CA, USA

    Loic Faye, John S. Greenwood, Eliot M. Herman, Arnd Sturm & Maarten J. Chrispeels

Authors
  1. Loic Faye
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John S. Greenwood
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eliot M. Herman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Arnd Sturm
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maarten J. Chrispeels
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Faye, L., Greenwood, J.S., Herman, E.M. et al. Transport and posttranslational processing of the vacuolar enzyme α-mannosidase in jack-bean cotyledons. Planta 174, 271–282 (1988). https://doi.org/10.1007/BF00394781

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00394781

Key words

Search

Navigation