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. 1976 Mar;57(3):430–436. doi: 10.1104/pp.57.3.430

Studies of Sulfate Utilization of Algae

15. Enzymes of Assimilatory Sulfate Reduction in Euglena and Their Cellular Localization 1

Christian Brunold a,2, Jerome A Schiff a,3
PMCID: PMC542040  PMID: 16659497

Abstract

Crude extracts of wild-type Euglena grown in the light (WTL) or in the dark (WTD) and a mutant lacking detectable plastid DNA (W3BUL) contain adenosine 5′-phosphosulfate (APS) sulfotransferase. Isotope dilution experiments indicate that adenosine 3′-phosphate 5′-phosphosulfate (PAPS) sulfotransferase is absent.

Thiosulfonate reductase, requiring addition of NADH or NADPH but not ferredoxin, and O-acetyl-l-serine sulfhydrylase, the two other enzymes of the bound intermediate pathway of assimilatory sulfate reduction, are also present. Increasing levels of all three enzymes were found in WTL, WTD, and W3BUL during logarithmic growth but the various activities were similar at comparable stages of growth in all three types of cell.

These results show that the three enzymes are not coded in the chloroplast DNA and are not restricted to Euglena cells having fully developed chloroplasts. Consistent with this, they do not increase during light-induced chloroplast development in resting cells and are found to be enriched in the mitochondrial fraction. Further resolution of this fraction on sucrose gradients shows that the APS sulfotransferase is associated with both the microbody (glyoxysomal) and mitochondrial fractions while the thiosulfonate reductase and O-acetyl-l-serine sulfhydrylase are associated only with the mitochondria. Thus the three known enzymes of the bound pathway of assimilatory sulfate reduction are present in Euglena mitochondria.

Although the activity of the entire bound pathway (APS to cysteine) is low in extracts, addition of dithiothreitol which releases free sulfite from the product of the APS sulfotransferase reaction, causes an increase in reduction activity indicating that a sulfite reductase is also present. It remains to be shown which reducing system is the significant one in vivo in Euglena.

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Selected References

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

  1. ABRAHAM A., BACHHAWATBK Effect of light and streptomycin on the incorporation of sulfate into sulfolipids in Euglena gracilis. Biochim Biophys Acta. 1963 Feb 19;70:104–106. doi: 10.1016/0006-3002(63)90729-7. [DOI] [PubMed] [Google Scholar]
  2. Becker M. A., Kredich N. M., Tomkins G. M. The purification and characterization of O-acetylserine sulfhydrylase-A from Salmonella typhimurium. J Biol Chem. 1969 May 10;244(9):2418–2427. [PubMed] [Google Scholar]
  3. Bovarnick J. G., Schiff J. A., Freedman Z., Egan J. M. Events surrounding the early development of Euglena chloroplasts: cellular origins of chloroplast enzymes in euglena. J Gen Microbiol. 1974 Jul;83(0):63–71. doi: 10.1099/00221287-83-1-63. [DOI] [PubMed] [Google Scholar]
  4. Cooper T. G., Beevers H. Mitochondria and glyoxysomes from castor bean endosperm. Enzyme constitutents and catalytic capacity. J Biol Chem. 1969 Jul 10;244(13):3507–3513. [PubMed] [Google Scholar]
  5. Davies W. H., Mercer E. I., Goodwin T. W. Some observations on the biosynthesis of the plant sulpholipid by Euglena gracilis. Biochem J. 1966 Feb;98(2):369–373. doi: 10.1042/bj0980369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ecklund P. R., Moore T. C. Correlations of Growth Rate and De-etiolation with Rate of Ent-Kaurene Biosynthesis in Pea (Pisum sativum L.). Plant Physiol. 1974 Jan;53(1):5–10. doi: 10.1104/pp.53.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gonzalez Porqué P., Baldesten A., Reichard P. The involvement of the thioredoxin system in the reduction of methionine sulfoxide and sulfate. J Biol Chem. 1970 May 10;245(9):2371–2374. [PubMed] [Google Scholar]
  8. Graves L. B., Jr, Trelease R. N., Grill A., Becker W. M. Localization of glyoxylate cycle enzymes in glyoxysomes in Euglena. J Protozool. 1972 Aug;19(3):527–532. doi: 10.1111/j.1550-7408.1972.tb03521.x. [DOI] [PubMed] [Google Scholar]
  9. Hilz H., Lipmann F. THE ENZYMATIC ACTIVATION OF SULFATE. Proc Natl Acad Sci U S A. 1955 Nov 15;41(11):880–890. doi: 10.1073/pnas.41.11.880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hodson R. C., Schiff J. A. Preparation of adenosine-3'-phosphate-51-phosphosulfate (PAPS): an improved enzymatic method using Chlorella pyrenoidosa. Arch Biochem Biophys. 1969 Jun;132(1):151–156. doi: 10.1016/0003-9861(69)90347-6. [DOI] [PubMed] [Google Scholar]
  11. Hodson R. C., Schiff J. A. Studies of Sulfate Utilization by Algae: 9. Fractionation of a Cell-free System from Chlorella into Two Activities Necessary for the Reduction of Adenosine 3'-Phosphate 5'-Phosphosulfate to Acid-Volatile Radioactivity. Plant Physiol. 1971 Feb;47(2):300–305. doi: 10.1104/pp.47.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. LYMAN H., EPSTEIN H. T., SCHIFF J. A. Studies of chloroplast development in Euglena. I. Inactivation of green colony formation by u.v. light. Biochim Biophys Acta. 1961 Jun 24;50:301–309. doi: 10.1016/0006-3002(61)90328-6. [DOI] [PubMed] [Google Scholar]
  14. Müller M., Hogg J. F., De Duve C. Distribution of tricarboxylic acid cycle enzymes and glyoxylate cycle enzymes between mitochondria and peroxisomes in Tetrahymena pyriformis. J Biol Chem. 1968 Oct 25;243(20):5385–5395. [PubMed] [Google Scholar]
  15. SHUSTER L., KAPLAN N. O. A specific b nucleotidase. J Biol Chem. 1953 Apr;201(2):535–546. [PubMed] [Google Scholar]
  16. Schiff J. A., Levinthal M. Studies of sulfate utilization by algae. 4. Properties of a cell-free sulfate-reducing system from chlorella. Plant Physiol. 1968 Apr;43(4):547–554. doi: 10.1104/pp.43.4.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schmidt A., Abrams W. R., Schiff J. A. Reduction of adenosine 5'-phosphosulfate to cysteine in extracts from Chlorella and mutants blocked for sulfate reduction. Eur J Biochem. 1974 Sep 16;47(3):423–434. doi: 10.1111/j.1432-1033.1974.tb03709.x. [DOI] [PubMed] [Google Scholar]
  18. Schmidt A. Sulfate reduction in a cell-free system of Chlorella. The ferredoxin dependent reduction of a protein-bound intermediate by a thiosulfonate reductase. Arch Mikrobiol. 1973 Oct 4;93(1):29–52. [PubMed] [Google Scholar]
  19. Schmidt A., Trebst A. The mechanism of photosynthetic sulfate reduction by isolated chloroplasts. Biochim Biophys Acta. 1969 Aug 5;180(3):529–535. doi: 10.1016/0005-2728(69)90031-0. [DOI] [PubMed] [Google Scholar]
  20. Stern A. I., Epstein H. T., Schiff J. A. Studies of Chloroplast Development in Euglena. VI. Light Intensity as a Controlling Factor in Development. Plant Physiol. 1964 Mar;39(2):226–231. doi: 10.1104/pp.39.2.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stern A. I., Schiff J. A., Epstein H. T. Studies of Chloroplast Development in Euglena. V. Pigment Biosynthesis, Photosynthetic Oxygen Evolution and Carbon Dioxide Fixation during Chloroplast Development. Plant Physiol. 1964 Mar;39(2):220–226. doi: 10.1104/pp.39.2.220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WILSON L. G., ASAHI T., BANDURSKI R. S. Yeast sulfate-reducing system. I. Reduction of sulfate to sulfite. J Biol Chem. 1961 Jun;236:1822–1829. [PubMed] [Google Scholar]
  23. Zeldin M. H., Schiff J. A. RNA metabolism during light-induced chloroplast development in euglena. Plant Physiol. 1967 Jul;42(7):922–932. doi: 10.1104/pp.42.7.922. [DOI] [PMC free article] [PubMed] [Google Scholar]

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