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. 1983;2(7):1035–1040. doi: 10.1002/j.1460-2075.1983.tb01542.x

Synthesis and processing of thionin precursors in developing endosperm from barley (Hordeum vulgare L.)

F Ponz 1, J Paz-Ares 1, C Hernández-Lucas 1, P Carbonero 1, F García-Olmedo 1,*
PMCID: PMC555231  PMID: 16453465

Abstract

Thionin is a lysine-rich polypeptide (mol. wt. 5000) which is synthesized in developing barley endosperm from ˜8 days to ˜30 days after anthesis. Two thionin precursors (THP1 and THP2) have been identified using monospecific antibodies (A-TH) prepared against the mature protein. THP1, which is the only polypeptide recognized in vitro by A-TH, is encoded by a 7.5S mRNA obtained from membrane-bound polysomes, and its alkylated derivative has an apparent mol. wt. of 17 800. THP2, which is selected together with mature thionin by A-TH among labelled proteins in vivo, differs from THP1 in apparent mol. wt. (17 400 alkylated) and in electrophoretic mobility at pH 3.2. Both THP1 and THP2 are competed out of the antigen-antibody complex by purified thionin. The conversion of THP2 into thionin, which has been demonstrated in a pulse-chase experiment in vivo, is a post-translational process. As it has not been possible to detect THP1 in vivo it is assumed that it is converted co-translationally into THP2. Final deposition of thionin as an extrinsic membrane protein, possibly associated with the endoplasmic reticulum, has been tentatively established on the basis of subcellular fractionation experiments.

Keywords: barley, endosperm, Hordeum vulgare L., protein biosynthesis, thionin

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  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blakesley R. W., Boezi J. A. A new staining technique for proteins in polyacrylamide gels using coomassie brilliant blue G250. Anal Biochem. 1977 Oct;82(2):580–582. doi: 10.1016/0003-2697(77)90197-x. [DOI] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Burr B., Burr F. A., Rubenstein I., Simon M. N. Purification and translation of zein messenger RNA from maize endosperm protein bodies. Proc Natl Acad Sci U S A. 1978 Feb;75(2):696–700. doi: 10.1073/pnas.75.2.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carbonero P., García-Olmedo F. Purothionins in Aegilops-Triticum spp. Experientia. 1969 Oct 15;25(10):1110–1111. doi: 10.1007/BF01901464. [DOI] [PubMed] [Google Scholar]
  6. Chrispeels M. J., Higgins T. J., Spencer D. Assembly of storage protein oligomers in the endoplasmic reticulum and processing of the polypeptides in the protein bodies of developing pea cotyledons. J Cell Biol. 1982 May;93(2):306–313. doi: 10.1083/jcb.93.2.306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Caleya R. F., Hernandez-Lucas C., Carbonero P., Garcia-Olmedo F. Gene expression in alloploids: genetic control of lipopurothionins in wheat. Genetics. 1976 Aug;83(4):687–699. doi: 10.1093/genetics/83.4.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dure L., Galau G. A. Developmental Biochemistry of Cottonseed Embryogenesis and Germination : XIII. REGULATION OF BIOSYNTHESIS OF PRINCIPAL STORAGE PROTEINS. Plant Physiol. 1981 Jul;68(1):187–194. doi: 10.1104/pp.68.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greene F. C. In Vitro Synthesis of Wheat (Triticum aestivum L.) Storage Proteins. Plant Physiol. 1981 Sep;68(3):778–783. doi: 10.1104/pp.68.3.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Larkins B. A., Hurkman W. J. Synthesis and deposition of zein in protein bodies of maize endosperm. Plant Physiol. 1978 Aug;62(2):256–263. doi: 10.1104/pp.62.2.256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Larkins B. A., Jones R. A., Tsai C. Y. Isolation and in vitro translation of zein messenger ribonucleic acid. Biochemistry. 1976 Dec 14;15(25):5506–5511. doi: 10.1021/bi00670a014. [DOI] [PubMed] [Google Scholar]
  12. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  13. Luthe D. S., Peterson D. M. Cell-free Synthesis of Globulin by Developing Oat (Avena sativa L.) Seeds. Plant Physiol. 1977 May;59(5):836–841. doi: 10.1104/pp.59.5.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mak A. S., Jones B. L. The amino acid sequence of wheat beta-purothionin. Can J Biochem. 1976 Oct;54(10):835–842. doi: 10.1139/o76-120. [DOI] [PubMed] [Google Scholar]
  15. Marcu K., Dudock B. Characterization of a highly efficient protein synthesizing system derived from commercial wheat germ. Nucleic Acids Res. 1974 Nov;1(11):1385–1397. doi: 10.1093/nar/1.11.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ohtani S., Okada T., Yoshizumi H., Kagamiyama H. Complete primary structures of two subunits of purothionin A, a lethal protein for brewer's yeast from wheat flour. J Biochem. 1977 Sep;82(3):753–767. doi: 10.1093/oxfordjournals.jbchem.a131752. [DOI] [PubMed] [Google Scholar]
  17. Okita T. W., Greene F. C. Wheat Storage Proteins : ISOLATION AND CHARACTERIZATION OF THE GLIADIN MESSENGER RNAs. Plant Physiol. 1982 Apr;69(4):834–839. doi: 10.1104/pp.69.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  19. Shapiro D. J., Taylor J. M., McKnight G. S., Palacios R., Gonzalez C., Kiely M. L., Schimke R. T. Isolation of hen oviduct ovalbumin and rat live albumin polysomes by indirect immunoprecipitation. J Biol Chem. 1974 Jun 25;249(12):3665–3671. [PubMed] [Google Scholar]
  20. Shapiro S. Z., Young J. R. An immunochemical method for mRNA purification. Application to messenger RNA encoding trypanosome variable surface antigen. J Biol Chem. 1981 Feb 25;256(4):1495–1498. [PubMed] [Google Scholar]
  21. Tai H. H., Chey W. Y. Development of radioimmunoassay for motilin. Anal Biochem. 1978 Jul 1;87(2):350–358. doi: 10.1016/0003-2697(78)90684-x. [DOI] [PubMed] [Google Scholar]
  22. Tolbert N. E. Isolation of subcellular organelles of metabolism on isopycnic sucrose gradients. Methods Enzymol. 1974;31:734–746. doi: 10.1016/0076-6879(74)31077-4. [DOI] [PubMed] [Google Scholar]
  23. Viotti A., Sala E., Marotta R., Alberi P., Balducci C., Soave C. Genes and mRNAs coding for zein polypeptides in Zea mays. Eur J Biochem. 1979 Dec;102(1):211–222. doi: 10.1111/j.1432-1033.1979.tb06282.x. [DOI] [PubMed] [Google Scholar]
  24. Wienand U., Feix G. Electrophoretic fractionation and translation in vitro of poly(rA)-containing RNA from maize endosperm. Evidence of two mRNAs coding for zein protein. Eur J Biochem. 1978 Dec;92(2):605–611. doi: 10.1111/j.1432-1033.1978.tb12783.x. [DOI] [PubMed] [Google Scholar]
  25. Yamagata H., Sugimoto T., Tanaka K., Kasai Z. Biosynthesis of storage proteins in developing rice seeds. Plant Physiol. 1982 Oct;70(4):1094–1100. doi: 10.1104/pp.70.4.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]

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