Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1977 May;59(5):836–841. doi: 10.1104/pp.59.5.836

Cell-free Synthesis of Globulin by Developing Oat (Avena sativa L.) Seeds 1

Dawn Sywassink Luthe a,2, David M Peterson a,3
PMCID: PMC543306  PMID: 16659952

Abstract

The primary storage protein synthesized during oat (Avena sativa L.) groat development is a globulin. Polysomes were isolated from oat groats 12 days after anthesis. These polysomes directed the incorporation of radioactive amino acids into protein in a cell-free protein synthesis system containing wheat germ supernatant. The Mg2+ optimum was 4 mm, the pH optimum was 6-8, and the amount of amino acid incorporation depended on polysome concentration. Incorporation of amino acids was linear for about 10 min and approached a maximum after 20 min. Using the initiation inhibitor, T-2 toxin, it was determined that about 36% of the amino acid incorporation was due to the initiation of new polypeptide chains. The in vitro product co-electrophoresed with authentic oat groat globulin on polyacrylamide-sodium dodecyl sulfate (SDS) gels. The cyanogen bromide peptides of the in vitro product partially corresponded with those from authentic globulin when electrophoresed on polyacrylamide-SDS gels. These data suggest that the in vitro product is primarily oat globulin. The polysome population was separated into membrane-bound and free polysomes. Membrane-bound polysomes synthesized about twice the amount of protein as did free polysomes. Products synthesized in vitro on both types of polysomes were essentially the same.

Full text

PDF
839

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beevers L., Poulson R. Protein Synthesis in Cotyledons of Pisum sativum L: I. Changes in Cell-Free Amino Acid Incorporation Capacity during Seed Development and Maturation. Plant Physiol. 1972 Apr;49(4):476–481. doi: 10.1104/pp.49.4.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burr B., Burr F. A. Zein synthesis in maize endosperm by polyribosomes attached to protein bodies. Proc Natl Acad Sci U S A. 1976 Feb;73(2):515–519. doi: 10.1073/pnas.73.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davies E., Larkins B. A., Knight R. H. Polyribosomes from peas: an improved method for their isolation in the absence of ribonuclease inhibitors. Plant Physiol. 1972 Nov;50(5):581–584. doi: 10.1104/pp.50.5.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goossens W., Préaux G., Lontie R. Cleavage of bovine serum albumin with cyanogen bromide and alignment of the fragments. Biochimie. 1973;55(10):1199–1207. doi: 10.1016/s0300-9084(74)80324-x. [DOI] [PubMed] [Google Scholar]
  5. Gray J. C., Kekwick R. G. The synthesis of the small subunit of ribulose 1,5-bisphosphate carboxylase in the french bean Phaseolus vulgaris. Eur J Biochem. 1974 May 15;44(2):491–500. doi: 10.1111/j.1432-1033.1974.tb03507.x. [DOI] [PubMed] [Google Scholar]
  6. Jones R. A., Larkins B. A., Tsai C. Y. Reduced synthesis of zein in vitro by a high lysine mutant of maize. Biochem Biophys Res Commun. 1976 Mar 22;69(2):404–410. doi: 10.1016/0006-291x(76)90536-2. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Larkins B. A., Bracker C. E., Tsai C. Y. Storage Protein Synthesis in Maize: Isolation of Zein-synthesizing Polyribosomes. Plant Physiol. 1976 May;57(5):740–745. doi: 10.1104/pp.57.5.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Larkins B. A., Dalby A. In vitro synthesis of zein-like protein by maize polyribosomes. Biochem Biophys Res Commun. 1975 Oct 6;66(3):1048–1054. doi: 10.1016/0006-291x(75)90746-9. [DOI] [PubMed] [Google Scholar]
  11. Marcus A., Efron D., Weeks D. P. The wheat embryo cell-free system. Methods Enzymol. 1974;30:749–754. doi: 10.1016/0076-6879(74)30073-0. [DOI] [PubMed] [Google Scholar]
  12. Morton R. K., Raison J. K. The separate incorporation of amino acids into storage and soluble proteins catalysed by two independent systems isolated from developing wheat endosperm. Biochem J. 1964 Jun;91(3):528–539. doi: 10.1042/bj0910528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. RABSON R., MANS R. J., NOVELLI G. D. Changes in cell-free amino acid incorporating activity during maturation of maize kernels. Arch Biochem Biophys. 1961 Jun;93:555–562. doi: 10.1016/s0003-9861(61)80052-0. [DOI] [PubMed] [Google Scholar]
  14. Smith K. E., Cannon M., Cundliffe E. Inhibition at the initiation level of eukaryotic protein synthesis by T-2 toxin. FEBS Lett. 1975 Jan 15;50(1):8–12. doi: 10.1016/0014-5793(75)81028-3. [DOI] [PubMed] [Google Scholar]
  15. Spande T. F., Witkop B., Degani Y., Patchornik A. Selective cleavage and modification of peptides and proteins. Adv Protein Chem. 1970;24:97–260. doi: 10.1016/s0065-3233(08)60242-9. [DOI] [PubMed] [Google Scholar]
  16. Stutz E., Noll H. Characterization of cytoplasmic and chloroplast polysomes in plants: evidence for three classes of ribosomal RNA in nature. Proc Natl Acad Sci U S A. 1967 Mar;57(3):774–781. doi: 10.1073/pnas.57.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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. 1975 Dec;56(6):780–785. doi: 10.1104/pp.56.6.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  19. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES