Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1994 Sep;106(1):119–125. doi: 10.1104/pp.106.1.119

Structure and Function of the Golgi Complex in Rice Cells (II. Purification and Characterization of Golgi Membrane-Bound Nucleoside Diphosphatase).

T Mitsui 1, M Honma 1, T Kondo 1, N Hashimoto 1, S Kimura 1, I Igaue 1
PMCID: PMC159506  PMID: 12232309

Abstract

Inosine diphosphatase bound to Golgi membranes was studied in rice (Oryza sativa L. cv Nipponkai) cells. The enzyme was solubilized with Triton X-100 from isolated rice Golgi membranes and was highly purified employing a series of chromatography steps in the presence of 20% glycerol and 0.1% Triton X-100. The apparent molecular mass of the enzyme was estimated by gel filtration column chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 200 and 55 kD, respectively. The isoelectric point of the enzyme was determined to be 7.5. The optimal pH for the enzyme activity was around 7 and the enzyme required Mg2+ for hydrolyzing activity. IDP, UDP, and GDP were effective substrate for the purified rice Golgi membrane-bound inosine diphosphatase, whereas activity with ADP, CDP, and thymidine 5[prime]-diphosphate was 10 to 20% of IDP. The Km values for IDP, UDP, and GDP were 0.48, 0.50, and 0.67 mM, respectively, and Vmax values were 1.85, 1.54, and 1.67 [mu]mol min-1 mg-1, respectively. These results indicate that the rice Golgi enzyme is a nucleoside diphosphatase that is specific for IDP, UDP, and GDP. Furthermore, this rice Golgi nucleoside diphosphatase stimulated the activity of glucan synthase I also localized in rice Golgi membranes. The results strongly support the view that this nucleoside diphosphatase is involved in regulation of [beta]-glucan synthesis in the plant Golgi complex.

Full Text

The Full Text of this article is available as a PDF (1,023.8 KB).

Selected References

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

  1. Abeijon C., Orlean P., Robbins P. W., Hirschberg C. B. Topography of glycosylation in yeast: characterization of GDPmannose transport and lumenal guanosine diphosphatase activities in Golgi-like vesicles. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6935–6939. doi: 10.1073/pnas.86.18.6935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dauwalder M., Whaley W. G., Kephart J. E. Phosphatases and differentiation of the Golgi apparatus. J Cell Sci. 1969 Mar;4(2):455–497. doi: 10.1242/jcs.4.2.455. [DOI] [PubMed] [Google Scholar]
  3. Doremus H. D., Blevins D. G. Purification and characterization of a specific nucleoside diphosphatase from soybean root nodules. Plant Physiol. 1988 May;87(1):41–45. doi: 10.1104/pp.87.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gardiner M., Chrispeels M. J. Involvement of the Golgi Apparatus in the Synthesis and Secretion of Hydroxyproline-rich Cell Wall Glycoproteins. Plant Physiol. 1975 Mar;55(3):536–541. doi: 10.1104/pp.55.3.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kuriyama Y. Studies on microsomal nucleoside diphosphatase of rat hepatocytes. Its purification, intramembranous localization, and turnover. J Biol Chem. 1972 May 25;247(10):2979–2988. [PubMed] [Google Scholar]
  6. 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]
  7. Leonard R. T., Hansen D., Hodges T. K. Membrane-bound Adenosine Triphosphatase Activities of Oat Roots. Plant Physiol. 1973 Apr;51(4):749–754. doi: 10.1104/pp.51.4.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. NOVIKOFF A. B., HEUS M. A microsomal nucleoside diphosphatase. J Biol Chem. 1963 Feb;238:710–716. [PubMed] [Google Scholar]
  9. Ray P. M. Regulation of beta-Glucan Synthetase Activity by Auxin in Pea Stem Tissue: I. Kinetic Aspects. Plant Physiol. 1973 Apr;51(4):601–608. doi: 10.1104/pp.51.4.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ray P. M., Shininger T. L., Ray M. M. ISOLATION OF beta-GLUCAN SYNTHETASE PARTICLES FROM PLANT CELLS AND IDENTIFICATION WITH GOLGI MEMBRANES. Proc Natl Acad Sci U S A. 1969 Oct;64(2):605–612. doi: 10.1073/pnas.64.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Staver M. J., Glick K., Baisted D. J. Uridine diphosphate glucose-sterol glucosyltransferase and nucleoside diphosphatase activities in etiolated pea seedlings. Biochem J. 1978 Feb 1;169(2):297–303. doi: 10.1042/bj1690297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Yamazaki M., Hayaishi O. Allosteric properties of nucleoside diphosphatase and its identity with thiamine pyrophosphatase. J Biol Chem. 1968 Jun 10;243(11):2934–2942. [PubMed] [Google Scholar]
  13. Yanagisawa K., Resnick D., Abeijon C., Robbins P. W., Hirschberg C. B. A guanosine diphosphatase enriched in Golgi vesicles of Saccharomyces cerevisiae. Purification and characterization. J Biol Chem. 1990 Nov 5;265(31):19351–19355. [PubMed] [Google Scholar]
  14. Yotsushima K., Mitsui T., Takaoka T., Hayakawa T., Igaue I. Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor). Plant Physiol. 1993 May;102(1):165–172. doi: 10.1104/pp.102.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES