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. 1996 Feb;110(2):621–629. doi: 10.1104/pp.110.2.621

The Superoxide Synthases of Plasma Membrane Preparations from Cultured Rose Cells.

T M Murphy 1, C K Auh 1
PMCID: PMC157758  PMID: 12226208

Abstract

Preparations of plasma membranes isolated from cultured rose (Rosa damascena Mill. cv Gloire de Guilan) cells synthesized O2- when incubated with either NADH or NADPH, as measured by an O2--specific assay based on the chemiluminescence of lucigenin. The activities were strongly dependent on the presence of Triton X-100. The Km for NADH was 159 [mu]M; that for NADPH was 19 [mu]M. Neither NADH- nor NADPH-dependent activity was inhibited by azide, an inhibitor of peroxidase, nor by antimycin A, an inhibitor of mitochondrial electron transport; both activities were inhibited by 30 to 100 nM diphenylene iodonium, an inhibitor of the mammalian NADPH oxidase. The NADH- and NADPH-dependent activities could be distinguished by detergent solubilization and ultracentrifugation: the NADH-dependent activity sedimented more easily, whereas the NADPH-dependent activity remained in suspension. One or both of these enzymes may provide the O2- seen when plant cells are exposed to pathogens or pathogen-associated elicitors; however, plasma membranes from rose cells treated with a Phytophthora elicitor had the same activity as control cells.

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Selected References

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

  1. Abo A., Boyhan A., West I., Thrasher A. J., Segal A. W. Reconstitution of neutrophil NADPH oxidase activity in the cell-free system by four components: p67-phox, p47-phox, p21rac1, and cytochrome b-245. J Biol Chem. 1992 Aug 25;267(24):16767–16770. [PubMed] [Google Scholar]
  2. Auh C. K., Murphy T. M. Plasma Membrane Redox Enzyme Is Involved in the Synthesis of O2- and H2O2 by Phytophthora Elicitor-Stimulated Rose Cells. Plant Physiol. 1995 Apr;107(4):1241–1247. doi: 10.1104/pp.107.4.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castillo F. J., Penel C., Greppin H. Peroxidase Release Induced by Ozone in Sedum album Leaves: Involvement of Ca. Plant Physiol. 1984 Apr;74(4):846–851. doi: 10.1104/pp.74.4.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Corbisier P., Houbion A., Remacle J. A new technique for highly sensitive detection of superoxide dismutase activity by chemiluminescence. Anal Biochem. 1987 Jul;164(1):240–247. doi: 10.1016/0003-2697(87)90392-7. [DOI] [PubMed] [Google Scholar]
  5. Cross A. R., Jones O. T. The effect of the inhibitor diphenylene iodonium on the superoxide-generating system of neutrophils. Specific labelling of a component polypeptide of the oxidase. Biochem J. 1986 Jul 1;237(1):111–116. doi: 10.1042/bj2370111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hodges T. K., Leonard R. T. Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol. 1974;32:392–406. doi: 10.1016/0076-6879(74)32039-3. [DOI] [PubMed] [Google Scholar]
  7. Hyland K., Voisin E., Banoun H., Auclair C. Superoxide dismutase assay using alkaline dimethylsulfoxide as superoxide anion-generating system. Anal Biochem. 1983 Dec;135(2):280–287. doi: 10.1016/0003-2697(83)90684-x. [DOI] [PubMed] [Google Scholar]
  8. Levine A., Tenhaken R., Dixon R., Lamb C. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell. 1994 Nov 18;79(4):583–593. doi: 10.1016/0092-8674(94)90544-4. [DOI] [PubMed] [Google Scholar]
  9. Morré D. J., Brightman A. O. NADH oxidase of plasma membranes. J Bioenerg Biomembr. 1991 Jun;23(3):469–489. doi: 10.1007/BF00771015. [DOI] [PubMed] [Google Scholar]
  10. Pou S., Pou W. S., Bredt D. S., Snyder S. H., Rosen G. M. Generation of superoxide by purified brain nitric oxide synthase. J Biol Chem. 1992 Dec 5;267(34):24173–24176. [PubMed] [Google Scholar]
  11. Price A. H., Taylor A., Ripley S. J., Griffiths A., Trewavas A. J., Knight M. R. Oxidative Signals in Tobacco Increase Cytosolic Calcium. Plant Cell. 1994 Sep;6(9):1301–1310. doi: 10.1105/tpc.6.9.1301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sandstrom R. P., Deboer A. H., Lomax T. L., Cleland R. E. Latency of Plasma Membrane H-ATPase in Vesicles Isolated by Aqueous Phase Partitioning : Increased substrate Accessibility or Enzyme Activation. Plant Physiol. 1987 Nov;85(3):693–698. doi: 10.1104/pp.85.3.693. [DOI] [PMC free article] [PubMed] [Google Scholar]

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