Abstract
Polyribosomes which have template activity in the wheat germ system have been isolated from developing pea seeds. Some of the translation products have identical mobilities to the vicilin and legumin subunits by SDS-PAGE. Certain products were specifically immunoprecipitated with antisera prepared against purified vicilin and legumin fractions. Various RNA fractions including poly A-rich RNA have also been isolated from polyribosomes and shown to direct the synthesis of polyripeptides whose properties are similar to the storage protein subunits. The results are discussed in relationship to other investigations with seed storage protein biosynthesis in vitro.
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Abbreviations
- DTT:
-
dithiothreitol
- SDS-PAGE:
-
SDS-polyacrylamide gel electrophoresis
- TCA:
-
tricarboxylic acid
References
Aviv, H., Leder, P.: Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc. Natl. Acad. Sci. USA 69, 1408–1412 (1972)
Bailey, C.J., Cobb, A., Boulter, D.: 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 (1970)
Beachy, R.B., Thompson, J.F., Madison, J.T.: Isolation of polyribosomes and messenger RNA active in in vitro synthesis of Soyabean seed proteins. Plant Physiol. 61, 139–144 (1978)
Beevers, L., Poulson, R.: Protein synthesis in cotyledons of Pisum sativum L. Plant Physiol. 49, 476–481 (1972)
Bjerrum, O.J., Ramlau, J., Clemmeson, I., Ingild, A., Bøg-Hansen, T.C.: An artefact in quantitative immunoelectrophoresis of spectrin caused by proteolytic activity in antibody preparations. Scand J. Immunol. 4, Suppl. 2, 81–88 (1975)
Bjerrum, O.J., Bøg-Hansen, T.C.: Immunochemical gel precipitation techniques for the analysis of membrane proteins In, “Membrane Methods” Maddy A.H., ed. London: Chapman and Hall 1977
Boime, I., Leder, P.: Protein synthesis directed by Eucephalomyocarditis virus in RNA III. Discrete polypeptides translated from a Monocistronic Messenger in vitro. Arch. Biochem. Biophys. 153, 706–713 (1972)
Bonner, W.M., Laskey, R.A.: A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88 (1974)
Brawerman, G.: The isolation of messenger RNA from mammalian cells. Methods in Enzymol. 30, Part F, 605–612 (1974)
Croy, R.R.D.: Ph.D. Thesis, University of Aberdeen, Localisation of the major proteins and Proteolytic enzymes in Phaseolus vulgaris L. at Germination 1977
Derbyshire, E., Wright, D.J., Boulter, D.: Legumin and vicilin, storage proteins of legume seeds. Phytochemistry 15, 3–24 (1976)
Evans, M.J., Lingrel, J.B. Hemoglobin mRNA synthesis of 9S and rRNA during erythroid cell development. Biochem J. 8, 3000–3005 (1969)
Fraenkel-Conrat, H.: Methods for investigating the essential groups for enzyme activity. Methods in Enzymology 4, Section I, 11, 251 (1957)
Gordon, M.E., Payne, P.I.: In vitro translation of the long-lived messenger ribonucleic acid of dry seeds. Plant 130, 269–273 (1976)
Harboe, N., Ingild, A.: Immunisation, isolation of immunoglobulins, Estimation of Antibody Titre. Scand. J. Immunol. 2, Supp 1, 161–164 (1973)
Higgins, T.J.V., Spencer, D.: Cell-free synthesis of pea seed proteins. Plant Physiol. 60, 655–661 (1977)
Krystosek, A., Cawthorn, M.L., Kabat, D.: Improved methods for purification and assay of eukaryotic messenger ribonucleic acids and ribosomes. J. Biol. Chem. 250, 6077–6084 (1975)
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage. T4. Nature 277, 680–685 (1970)
Larkins, B.A., Bracker, C.E., Tsai, C.Y.: Storage protein synthesis in maize. Plant Physiol. 57, 740–745 (1976a)
Larkins, B.A., Jones, R.A., Tsai, C.Y.: Isolation and in vitro translation of Zein Messenger Ribonucleic Acid. Biochemstry 15, 5506–5510 (1976b)
Larkins, B.A., Davis, E.: Polyribosomes from peas. An attempt to characterise the total, free and membrane-bound polysomal population. Plant Physiol. 55, 749–756 (1975)
Luthe, D.S., Peterson, D.M.: Cell-free synthesis of globulin by developing oat (Avena sativa L.) seeds. Plant Physiol. 59, 836–841 (1977)
Millerd, A.: Biochemistry of legume seed proteins. Ann. Rev. Plant Physiol. 26, 53–72 (1975)
Millerd A, Spencer, D.: Changes in RNA synthesising activity and template activity in nucleic from cotyledons of developing pea seeds, Aust. J. Plant Physiol. 1, 33–1341 (1974)
Muntz, K.: Cell Specialisation Processes during biosynthesis and storage of proteins in plant seeds in ‘Regulation of developmental processes in plants’. p. 70–97. Schutte H.R., Gross, D., eds. Jena: VEB Gustav Fischer 1978
Pelham, H.B.R., Jackson, R.J.: An efficient mRNA-dependent translation system for reticulocyte lysates. Eur. J. Biochem. 67, 247–256 (1976)
Pusztai, A., Watt, W.B.: Glycoprotein II. The isolation and characterisation of a major antigenic and non-haemagglutinating Glycoprotein from Phaseolus vulgaris L. Biochim. Biophys. Acta. 207, 413–431 (1970)
Raacke, I.D.: Protein synthesis in ripening pea seeds. J. Analysis of whole seeds. Biochem. J. 66, 101–110 (1957)
Scholtz, G., Richter, J., Manteuffel, R.: Studies on seed globulins from legumes I. separation and purification of legumin and vicilin from V. faba L. by zone precipitation. Biochem. Physiol. Pflanzen 166, 163–172 (1974)
Sun, S.M., Buchbinder, B.U., Hall, T.C.: Cell-free synthesis of the major storage protein of the bean Phaseolus vulgaris L. Plant Physiol. 56, 780–785 (1975)
Wright, D.J., Boulter, D.: Purification and subunit structure of legumin of Vicia faba L. (Broad bean). Biochem. J. 141, 413–418 (1974)
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Marta Evans, I., Croy, R.R.D., Hutchinson, P. et al. Cell free synthesis of some storage protein subunits by polyribosomes and RNA isolated from developing seeds of pea (Pisum sativum L.). Planta 144, 455–462 (1979). https://doi.org/10.1007/BF00380122
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DOI: https://doi.org/10.1007/BF00380122