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. 1984 Sep;76(1):219–223. doi: 10.1104/pp.76.1.219

Subcellular Localization of Spermidine Synthase in the Protoplasts of Chinese Cabbage Leaves 1

Ram K Sindhu 1, Seymour S Cohen 1
PMCID: PMC1064259  PMID: 16663802

Abstract

Previous studies on the presence of spermidine synthase (EC 2.5.1.16) in the protoplasts of Chinese cabbage (Brassica pekinensis var Pak Choy) leaves had detected a small but significant fraction of the enzyme in a crude chloroplast fraction (Cohen, Balint, Sindhu 1981 Plant Physiol 68: 1150-1155). To establish whether this enzyme is truly a chloroplast component, we have isolated purified intact chloroplasts from protoplasts by density gradient centrifugation in silica sols (Ludox AM). Such chloroplasts contained all of the diaminopimelate decarboxylase (EC 4.1.1.20) of the protoplasts, but were essentially devoid of spermidine synthase. Control experiments showed that the latter had not been inactivated under conditions of isolation, purification, and assay of the intact chloroplasts. Isolation and assay of protoplast vacuoles in a further examination of the supernatant fluid containing the enzyme revealed a significant fraction of the enzyme in the vacuole fraction. However this fraction was found to contain similar proportions of a soluble enzyme, glucose 6-phosphate dehydrogenase. It has been concluded that vacuolar fractions are difficultly separable from soluble cytoplasmic material, which is probably the only compartment containing spermidine synthase.

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Selected References

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

  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beer S. V., Kosuge T. Spermidine and spermine--polyamine components of turnip yellow mosaic virus. Virology. 1970 Apr;40(4):930–938. doi: 10.1016/0042-6822(70)90139-x. [DOI] [PubMed] [Google Scholar]
  3. Boller T., Kende H. Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 1979 Jun;63(6):1123–1132. doi: 10.1104/pp.63.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen S. S., Balint R., Sindhu R. K. The synthesis of polyamines from methionine in intact and disrupted leaf protoplasts of virus-infected chinese cabbage. Plant Physiol. 1981 Nov;68(5):1150–1155. doi: 10.1104/pp.68.5.1150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. S., Greenberg M. L. Spermidine, an intrinsic component of turnip yellow mosaic virus. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5470–5474. doi: 10.1073/pnas.78.9.5470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. S., McCormick F. P. Polyamines and virus multiplication. Adv Virus Res. 1979;24:331–387. doi: 10.1016/s0065-3527(08)60397-8. [DOI] [PubMed] [Google Scholar]
  7. Cohen S. S. Mitochondria and chloroplasts revisited. Am Sci. 1973 Jul-Aug;61(4):437–445. [PubMed] [Google Scholar]
  8. Guarino L. A., Cohen S. S. The estimation of turnover of spermidine in Anacystis nidulans. Anal Biochem. 1979 May;95(1):73–76. doi: 10.1016/0003-2697(79)90186-6. [DOI] [PubMed] [Google Scholar]
  9. Hartmann T., Schoofs G., Wink M. A chloroplast-localized lysine decarboxylase of Lupinus polyphyllus: the first enzyme in the biosynthetic pathway of quinolizidine alkaloids. FEBS Lett. 1980 Jun 16;115(1):35–38. doi: 10.1016/0014-5793(80)80721-6. [DOI] [PubMed] [Google Scholar]
  10. Heber U., Pon N. G., Heber M. Localization of Carboxydismutase & Triosephosphate Dehydrogenases in Chloroplasts. Plant Physiol. 1963 May;38(3):355–360. doi: 10.1104/pp.38.3.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. JOHNSON M. W., MARKHAM R. Nature of the polyamine in plant viruses. Virology. 1962 Jun;17:276–281. doi: 10.1016/0042-6822(62)90117-4. [DOI] [PubMed] [Google Scholar]
  12. Leipold B. Effect of spermidine on the RNA-A protein complex isolated from the RNA bacteriophage MS2. J Virol. 1977 Feb;21(2):445–450. doi: 10.1128/jvi.21.2.445-450.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Price C. A., Dowling E. L. On the purification of the silica sol Ludox AM. Anal Biochem. 1977 Sep;82(1):243–245. doi: 10.1016/0003-2697(77)90155-5. [DOI] [PubMed] [Google Scholar]
  14. Ramakrishna S., Guarino L., Cohen S. S. Polyamines of Anacystis nidulans and metabolism of exogenous spermidine and spermine. J Bacteriol. 1978 Jun;134(3):744–750. doi: 10.1128/jb.134.3.744-750.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Shah S. P.J., Cossins E. A. Pteroylglutamates and methionine biosynthesis in isolated chloroplasts. FEBS Lett. 1970 Apr 16;7(3):267–270. doi: 10.1016/0014-5793(70)80177-6. [DOI] [PubMed] [Google Scholar]
  16. Sindhu R. K., Cohen S. S. Propylamine transferases in chinese cabbage leaves. Plant Physiol. 1984 Mar;74(3):645–649. doi: 10.1104/pp.74.3.645. [DOI] [PMC free article] [PubMed] [Google Scholar]

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