Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1986 Apr;80(4):950–955. doi: 10.1104/pp.80.4.950

Glycoproteins in the Matrix of Glyoxysomes in Endosperm of Castor Bean Seedlings 1

Elma González 1
PMCID: PMC1075235  PMID: 16664747

Abstract

The matrix of glyoxysomes from endosperm of castor bean (Ricinus communis cv Hale) seedlings has been analyzed for the presence of glycosylated proteins. Glyoxysome preparations were monitored for organelle homogeneity by electron microscopy and enzyme marker activities. Glyoxysomes were essentially free of endoplasmic reticulum, mitochondria, and protein bodies. At least eight glyoxysomal matrix glycopeptides ranging in size from 39 to 160 kilodaltons were identified by their affinity for concanavalin A. The glyoxysomal glycoproteins were shown to be radioactively labeled when endosperm was allowed to incorporate glucosamine. Incorporation of glucosamine was inhibited by tunicamycin under conditions which did not inhibit protein synthesis. Hydrolysis of glyoxysomal extracts and subsequent analysis by paper chromatography showed that the labeled precursor was incorporated into the glycoprotein without prior dispersion of the label into amino acids. The present data demonstrate the occurrence of N-linked, high mannose oligosaccharides on polypeptides of the glyoxysomal matrix. This finding is discussed in relation to pathways of protein maturation and transport during glyoxysomal biogenesis.

Full text

PDF
950

Images in this article

952Fig. 1
on p.952
952Fig. 2
on p.952
952Fig. 3
on p.952
953Fig. 4
on p.953
954Fig. 6
on p.954

Selected References

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

  1. 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]
  2. Bowden L., Lord J. M. Similarities in the polypeptide composition of glyoxysomal and endoplasmic-reticulum membranes from castor-bean endosperm. Biochem J. 1976 Feb 15;154(2):491–499. doi: 10.1042/bj1540491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burridge K. Direct identification of specific glycoproteins and antigens in sodium dodecyl sulfate gels. Methods Enzymol. 1978;50:54–64. doi: 10.1016/0076-6879(78)50007-4. [DOI] [PubMed] [Google Scholar]
  4. Conder M. J., Lord J. M. Heterogeneous distribution of glycosyltransferases in the endoplasmic reticulum of castor bean endosperm. Plant Physiol. 1983 Jun;72(2):547–552. doi: 10.1104/pp.72.2.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Donaldson R. P., Beevers H. Lipid composition of organelles from germinating castor bean endosperm. Plant Physiol. 1977 Feb;59(2):259–263. doi: 10.1104/pp.59.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frevert J., Kindl H. Plant microbody proteins. Purification and glycoprotein nature of glyoxysomal isocitrate lyase from cucumber cotyledons. Eur J Biochem. 1978 Dec 1;92(1):35–43. doi: 10.1111/j.1432-1033.1978.tb12720.x. [DOI] [PubMed] [Google Scholar]
  7. Gietl C., Hock B. Organelle-bound malate dehydrogenase isoenzymes are synthesized as higher molecular weight precursors. Plant Physiol. 1982 Aug;70(2):483–487. doi: 10.1104/pp.70.2.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gonzalez E., Beevers H. Role of the endoplasmic reticulum in glyoxysome formation in castor bean endosperm. Plant Physiol. 1976 Mar;57(3):406–409. doi: 10.1104/pp.57.3.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. González E. Aggregated Forms of Malate and Citrate Synthase are Localized in Endoplasmic Reticulum of Endosperm of Germinating Castor Bean. Plant Physiol. 1982 Jan;69(1):83–87. doi: 10.1104/pp.69.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hori H., Elbein A. D. Tunicamycin inhibits protein glycosylation in suspension cultured soybean cells. Plant Physiol. 1981 May;67(5):882–886. doi: 10.1104/pp.67.5.882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang A. H., Beevers H. Localization of enzymes within microbodies. J Cell Biol. 1973 Aug;58(2):379–389. doi: 10.1083/jcb.58.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kagawa T. Organelle-specific isozymes of citrate synthase in the endosperm of developing ricinus seedlings. Plant Physiol. 1981 Oct;68(4):845–850. doi: 10.1104/pp.68.4.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kindl H., Köller W., Frevert J. Cytosolic precursor pools during glyoxysome biosynthesis. Hoppe Seylers Z Physiol Chem. 1980;361(3):465–467. [PubMed] [Google Scholar]
  14. 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]
  15. Lord J. M., Bowden L. Evidence that glyoxysomal malate synthase is segregated by the endoplasmic reticulum. Plant Physiol. 1978 Feb;61(2):266–270. doi: 10.1104/pp.61.2.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lord J. M., Kagawa T., Moore T. S., Beevers H. Endoplasmic reticulum as the site of lecithin formation in castor bean endosperm. J Cell Biol. 1973 Jun;57(3):659–667. doi: 10.1083/jcb.57.3.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Markwell M. A. A new solid-state reagent to iodinate proteins. I. Conditions for the efficient labeling of antiserum. Anal Biochem. 1982 Sep 15;125(2):427–432. doi: 10.1016/0003-2697(82)90025-2. [DOI] [PubMed] [Google Scholar]
  18. Marriott K. M., Tanner W. Dolichylphosphate-dependent Glycosyl Transfer Reactions in the Endoplasmic Reticulum of Castor Bean Endosperm. Plant Physiol. 1979 Sep;64(3):445–449. doi: 10.1104/pp.64.3.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mellor R. B., Krusius T., Lord J. M. Analysis of glycoconjugate saccharides in organelles isolated from castor bean endosperm. Plant Physiol. 1980 Jun;65(6):1073–1075. doi: 10.1104/pp.65.6.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mellor R. B., Roberts L. M., Lord J. M. Glycosylation of exogenous protein by endoplasmic-reticulum membranes from castor-bean (Ricinus communis) endosperm. Biochem J. 1979 Aug 15;182(2):629–631. doi: 10.1042/bj1820629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Riezman H., Weir E. M., Leaver C. J., Titus D. E., Becker W. M. Regulation of Glyoxysomal Enzymes during Germination of Cucumber: 3. IN VITRO TRANSLATION AND CHARACTERIZATION OF FOUR GLYOXYSOMAL ENZYMES. Plant Physiol. 1980 Jan;65(1):40–46. doi: 10.1104/pp.65.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roberts L. M., Lord J. M. Synthesis and posttranslational segregation of glyoxysomal isocitrate lyase from castor bean endosperm. Eur J Biochem. 1981 Sep;119(1):43–49. doi: 10.1111/j.1432-1033.1981.tb05574.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES