Skip to main content
Springer Nature Link
Log in

The Golgi apparatus mediates the transport of phytohemagglutinin to the protein bodies in bean cotyledons

  • Published:
Planta Aims and scope Submit manuscript

Abstract

When developing cotyledons of Phaseolus vulgaris L. were labeled with [3H]fucose, fucose-labeled phytohemagglutinin (PHA) was found in organelles with average densities of 1.13 g cm-3 and 1.22 g cm-3. The position of these organelles on isopycnic sucrose gradients was independent of the presence of MgCl2 and ethylenediaminetetraacetate in the media, indicating that the fucose-labeled PHA was not associated with the rough endoplasmic reticulum (ER). The organelles with a density of 1.13 g cm-3 were identified as membranes of the Golgi apparatus on the basis of the similarity of their sedimentation properties and those of the Golgi marker enzyme, inosine diphosphatase, in both isopycnic and rate-zonal sucrose gradients. The organelles with a density of 1.22 g cm-3 were identified as small (0.1–0.4 μm), electron-dense vesicles with a protein content similar to that of the protein bodies. Pulsechase experiments with [3H]fucose indicated that fucose-labeled PHA first appeared in the Golgi-apparatus-derived membranes and later in the dense vesicles. Fucose-labeled PHA chased out of the Golgi apparatus first, then out of the dense vesicles, and accumulated in the soluble portion of the homogenate which contained the contents of the broken protein bodies. Fucose-labeled PHA chased out of the two types of organelles with a t 1/2 of 20–30 min, a rate three to four times faster than newly synthesized PHA chases out of the bulk of the ER (Chrispeels, M.J., Bollini, R., 1982, Plant Physiol. 70, 1425–1428). This result indicates that the Golgi apparatus is a much smaller compartment than the ER in the storage parenchyma cells. The sodium ionophore, monensin, which interferes with the function of the Golgi apparatus of animal cells, blocks the biosynthesis and—or transport of fucose- and galactose-labeled macromolecules to the cotyledon cell walls. Monensin also blocks the transport of labeled PHA out of the Golgi apparatus and into the protein bodies. These results provide the first biochemical evidence that a specific storage protein which accumulates in seeds is modified in, and passes through, the Golgi apparatus on its way to the protein bodies.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

endo-H:

endo-β-N-acetylglucosaminidase

EDTA:

ethylenediaminetetraacetate

ER:

endoplasmic reticulum

IDPase:

inosinediphosphatase

PHA:

phytohemagglutinin

SDS-PAGE:

sodium dodecylsulfate polyacrylamide-gel electrophoresis

References

  • 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

    Google Scholar 

  • Bailey, C.J., Cobb, A., Boulter, D. (1970) A cotyledon slice system for the autoradiographic study of the synthesis and intracellular transport of the seed storage proteins of Vicia faba. Planta 95, 103–118

    Google Scholar 

  • Barker, R.D.J., Derbyshire, E., Yarwood, A., Boulter, D. (1976) Purification and characterization of the major storage proteins of Phaseolus vulgaris seeds and their intracellular and cotyledonary distribution. Phytochemistry 15, 751–757

    Google Scholar 

  • Baumgartner, B., Tokuyasu, K.T., Chrispeels, M.J. (1980) Immunocytochemical localization of phaseolin in the endoplasmic reticulum of developing bean (Phaseolus vulgaris) cotyledons. Planta 150, 419–425

    Google Scholar 

  • Beevers, L., Mense, R.M. (1977) Glycoprotein biosynthesis in cotyledons of Pisum sativum L. Involvement of lipid-linked intermediates. Plant Physiol. 60, 703–708

    Google Scholar 

  • 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

    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., 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

    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 for tritium-labeled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Chrispeels, M.J., Higgins, T.J.V., Craig, S., Spencer, D. (1982) The 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 

  • Dauwalder, M., Whaley, W.G., Kephart, J.E. (1969) Phosphatases and differentiation of the Golgi apparatus. J. Cell Sci 4, 455–497

    Google Scholar 

  • 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

    Google Scholar 

  • Delmer, D.P., Kulow, C., Ericson, M.C. (1978) Glycoprotein synthesis in plants II. Structure of the mannolipid intermediate. Plant Physiol. 61, 25–29

    Google Scholar 

  • Dieckert, J.W., Dieckert, M.C. (1976) The chemistry and cell biology of the vacuolar proteins of seeds. J. Food Sci. 41, 475–482

    Google Scholar 

  • Farquhar, M.G., Palade, G.E. (1981) The Golgi apparatus (complex) — (1954–1981) — from artifact to center stage. J. Cell Biol. 91, 77s-103s

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • Gardiner, M., Chrispeels, M.J. (1975) Involvement of the Golgi apparatus in the synthesis and secretion of hydroxyprolinerich cell wall glycoproteins. Plant Physiol. 55, 536–541

    Google Scholar 

  • Grimes, H., Boss, W.F., Morre, D.J., Mollenhauer, H.M. (1982) Ultrastructural observations of embryogenic suspension culture cells of carrot following exposure to monensin. (Abstr.) Plant Physiol. 69, Suppl., 48

    Google Scholar 

  • Harris, N., (1979) Endoplasmic reticulum in developing seeds of Vicia faba. Planta 146, 63–69

    Google Scholar 

  • Harris, N., Oparka, K.J. (1983) Connections between dictyosomes, ER and GERL in cotyledons of mung bean (Vigna radiata L.). Protoplasma 114, 93–102

    Google Scholar 

  • Higgins, T.J.V., Spencer, D. (1981) Precursor forms of pea vicilin subunits. Modification by microsomal membranes during cell-free translation. Plant Physiol. 65, 205–211

    Google Scholar 

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

    Google Scholar 

  • Hurkman, W.J., Beevers, L. (1982) Sequestration of pea reserve proteins by rough microsomes. Plant Physiol. 69, 1414–1417

    Google Scholar 

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

    Google Scholar 

  • Lord, J.M., Kagawa, T., Moore, T.S., Beevers, H. (1973) Endoplasmic reticulum as the site of lecithin formation in castor bean endosperm. J. Cell Biol. 57, 659–667

    Google Scholar 

  • 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. Vég. 11, 519–537

    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 

  • Munro, J.R., Narasimhan, S., Wetmore, S., Riordan, J.R., Schachter, H. (1975) Intracellular localization of GDP-L-fucose glycoprotein and CMP-sialic acid apolipoprotein glycososyltransferases in rat and pork liver. Arch. Biochem. Biophys. 169, 269–277

    Google Scholar 

  • Nishimoto, S.K., Kajiwara, T., Ledger, P.W., Tanzer, M.L. (1982) Effects of the ionophore monensin on type II collagen and proteoglycan synthesis and secretion by cultured chondrocytes. J. Biol. Chem. 257, 11712–11716

    Google Scholar 

  • Ohnishi, T., Gall, R.S., Mayer, M.L. (1975) An improved assay at inorganic phosphate in the presence of extractable phosphate compounds: application to the ATPase assay in the presence of phosphocreatine. Anal. Biochem. 69, 261–267

    Google Scholar 

  • Ray, P.M. (1980) Cooperative action of β-glucan synthetase and UDP-xylose xylosyltransferase of Golgi membranes in the synthesis of xyloglucan-like polysaccharides. Biochim. Biophys. Acta 629, 431–444

    Google Scholar 

  • Ray, P.M., Eisinger, R., Robinson, D.G. (1976) Organelles involved in cell wall polysaccharide formation and transport in pea cells. Ber. Dtsch. Bot. Ges. 89, 129–146

    Google Scholar 

  • Ray, P.M., Shininger, T.L., Ray, M.M. (1969) Isolation of β-glucan synthetase particles from plant cells and identification with Golgi membranes. Proc. Natl. Acad. Sci. USA 64, 605–612

    Google Scholar 

  • Sengupta, C., Deluca, V., Bailey, D.S., Verma, D.P.S. (1981) Post-translational processing of 7S and 11S components of soybean storage proteins. Plant Mol. Biol. 1, 19–34

    Google Scholar 

  • Sottocasa, G.L., Kuylenstierna, B., Ernster, L., Bergstrand, A. (1967) An electron-transport system associated with the outer membrane of liver mitochondria. J. Cell Biol. 32, 415–438

    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 

  • Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31–43

    Google Scholar 

  • 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

    Google Scholar 

  • Sun, S.M., Mutschler, M.A., Bliss, F.A., Hall, T.C. (1978) Protein synthesis and accumulation in bean cotyledons during growth. Plant Physiol. 61, 918–923

    Google Scholar 

  • Tartakoff, A., Vassalli, P. (1978) Comparative studies of intracellular transport of secretory proteins. J. Cell Biol. 79, 694–707

    Google Scholar 

  • Uchida, N., Smilowitz, H., Ledger, P.W., Tanzer, M.L. (1980) Kinetic studies of the intracellular transport of procollagen and fibronectin in human fibroblasts. Effects of the monovalent ionophore monensin. J. Biol. Chem. 25, 8638–8644

    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 

  • Vladutiu, G.D., and Rattazzi, M.C. (1980) The effect of monesin on β-hexosaminidase transport in normal and I-cell fibroblasts. Biochem. J. 192, 813–820

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, C-016, University of California-San Diego, 92093, La Jolla, CA, USA

    Maarten J. Chrispeels

Authors
  1. 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

Chrispeels, M.J. The Golgi apparatus mediates the transport of phytohemagglutinin to the protein bodies in bean cotyledons. Planta 158, 140–151 (1983). https://doi.org/10.1007/BF00397707

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00397707

Key words

Search

Navigation