Abstract
Endosperm tissue was excised from Ricinus communis plants at different stages during seed maturation. The various stages were characterized on the basis of total RNA, protein and lipid content. Polyadenylated RNA was recovered from the total RNA by affinity chromatography on oligo(dT)cellulose. With the exception of that isolated from dry seeds, this poly(A+) RNA actively programmed protein synthesis in cell-free systems containing either wheat germ S30 extracts or nuclease treated rabbit reticulocyte lysates at each developmental stage examined. Translational products were separated electrophoretically and were visualized by fluorography. The capacity to synthesize protein was also estimated during ‘in vivo’ labelling studies. Developmental changes in the capacity of maturing endosperm tissue to synthesize a characteristic protein, R. communis agglutinin, were followed by immunoprecipitating this protein from the total ‘in vitro’ products synthesized at various stages. Endoplasmic reticulum membranes were isolated from maturing endosperm tissue by sucrose density gradient centrifugation. The role of the endoplasmic reticulum (ER) in protein glycosylation was indicated by (a) localizing the enzymes catalysing the incorporation of N-acetylglucosamine and mannose into mono- and oligosaccharide lipid and into glycoprotein, (b) localizing particulate 3H-labelled glycoprotein amongst cellular fractions prepared from endosperm tissue which had been incubated with [3H]N-acetylglucosamine.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- polyCA+)RNA:
-
polyadenylated RNA
- SDS:
-
sodiumdodecylsulphate
- TCA:
-
trichloroacetic acid
- RCA120 :
-
Ricinus communis agglutinin, type I
- NP40:
-
Nonidet P40
- PMSF:
-
phenylmethylsulphonyl fluoride
References
Bailey, C.J., Cobb, A., Boulter, D. (1970) A cotyledon slice system for the electron autoradiographic study of the synthesis and intracellular transport of the seed storage protein of Vicia faba. Planta 95, 103–118
Beevers, H. (1961) Metabolic production of sucrose from fat. Nature (London) 191, 433–436
Reevers, H. (1975) Organelles from castor bean seedlings: biochemical role in gluconeogenesis and phospholipid biosynthesis. In: Recent advances in the chemistry and biochemistry of plant lipids pp. 287–299. Galliard, T., Mercer, E.I. eds. Academic Press, London
Beevers, H. (1979) Microbodies from higher plants. Annu. Rev. Plant Physiol. 30, 159–193
Bollini, R., Chrispeels, M.J. (1979) The rough endoplasmic reticulum is the site of reserve protein synthesis in developing Phaseolus vulgaris. Planta 146, 487–501
Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium labeled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88
Bray, G.A. (1960) A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Anal. Biochem. 1, 270–285
Burr, B., Burr, F.A. (1976) Zein synthesis in maize endosperm by polyribosomes attached to protein bodies. Proc. Natl. Acad. Sci. USA 73, 515–519
Czicki, U., Lennarz, W.J. (1977) Localization of the enzyme system for the glycosylation of proteins via the lipid-linked pathway in rough endoplasmic reticulum. J. Biol. Chem. 252, 7901–7904
Kiely, M.L., McKnight, G.S., Schimke, R.T. (1976) Studies on the attachment of carbohydrate to ovalbumin nascent chains in hen oviduct. J. Biol. Chem. 251, 5490–5495
Kornfeld, R., Kornfeld, S. (1976) Comparative aspects of glycoprotein structure. Annu. Rev. Biochem. 45, 217–237
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685
Larkins, B.A., Hurkman, W.J. (1978) Synthesis and deposition of zein in protein bodies of maize endosperm. Plant Physiol. 62, 256–263
Lord, J.M. (1976) Phospholipid synthesis and exchange in castor bean endosperm homogenates. Plant Physiol. 57, 218–223
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
Lowry, O.H., Rosebrough, M.J., Farr, A.L., Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275
Mans, R.J., Novelli, G.D. (1961) Measurement of the incorporation of radioactive amino acids into protein by a filter-paper disc method. Arch. Biochem. Biophys. 94, 48–53
Mazliak, P., Douady, D., Demandre, C., Kader, J.C. (1975) Exchange processes between organelles involved in membrane lipid biosynthesis. In: Recent advances in the chemistry and biochemistry of plant lipids. pp. 301–318. Galhard, T., Mercer, E.I. eds. Academic Press, London
Mellor, R.B., Lord, J.M. (1979) Formation of lipid-linked mono-and oligosaccharides from GDP-mannose by castor bean endosperm homogenates. Planta 146, 91–99
Mellor, R.B., Roberts, L.M., Lord, J.M. (1979) Glycosylation of exogenous proteins by endoplasmic reticulum membranes from castor bean (Ricinus communis) endosperm. Biochem. J. 182, 629–631
Mettler, I.J., Beevers, H. (1979) Isolation and characterization of the protein body membrane of castor beans. Plant Physiol. 64, 506–511
Nicholson, G.L., Blanstein, J., Etzler, M.E. (1974) Characterization of two plant lectins from Ricinus communis and their quantitative interaction with a murine lymphoma. Biochemistry 13, 196–204
Olsnes, S., Refsnes, K., Christensen, T.B., Pihl, A. (1975) Studies on the structure and properties of lectins from Abrus precatorius and Ricinus communis. Biochim. Biophys. Acta 405, 1–10
Parodi, A.J., Leloir, L.F. (1979) The role of lipid intermediates in the glycosylation of proteins in the eukaryotic cell. Biochim. Biophys. Acta 559, 1–37
Pelham, H.R.B., Jackson, R.J. (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. 67, 247–256
Püchel, M., Müntz, K., Parthier, B., Aurich, O., Bassüner, R., Mantenfel, R., Schmidt, P. (1979) RNA metabolism and membrane bound polysomes in relation to globin synthesis in cotyledons of developing field bean (Vicia faba L.). Eur. J. Biochem. 96, 321–329
Roberts, L.M., Lord, J.M. (1979a) Ribonucleic acid synthesis in germinating castor bean endosperm. J. Exp. Bot. 30, 739–749
Roberts, L.M., Lord, J.M. (1979b) Developmental changes in the activity of messenger RNA isolated from germinating castor bean endosperm. Plant Physiol. 64, 630–634
Roberts, B.E., Patterson, B.M. (1973) Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cellfree system from commercial wheat germ. Proc. Natl. Acad. Sci. USA, 70, 2330–2334
Tulley, R.E., Beevers, H. (1976) Protein bodies of castor bean endosperm. Isolation, fractionation, and characterization of protein components. Plant Physiol. 58, 710–716
Wirtz, K.W.A. (1974) Transfer of phospholipids between membranes. Biochim. Biophys. Acta 344, 95–117
Youle, R.J., Huang, A.H.C. (1976) Protein bodies from the endosperm of castor bean. Subfractionation, protein components, lectins, and changes during germination. Plant Physiol. 58, 703–709
Youle, R.J., Huang, A.H.C. (1978) Albumin storage proteins in the protein bodies of castor bean. Plant Physiol. 61, 13–16
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Roberts, L.M., Lord, J.M. Protein biosynthetic capacity in the endosperm tissue of ripening castor bean seeds. Planta 152, 420–427 (1981). https://doi.org/10.1007/BF00385358
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00385358