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. 1980 Aug;66(2):291–294. doi: 10.1104/pp.66.2.291

Preparation of Chloroplasts from Euglena Highly Active in Protein Synthesis 1

William Ortiz 2,3, Ellen M Reardon 2, Carl A Price 2
PMCID: PMC440585  PMID: 16661424

Abstract

Chloroplasts can be obtained by gentle lysis or mild shear of spheroplasts of vitamin B12-deficient Euglena gracilis and then purified by isopycnic sedimentation on gradients of Ludox AM or Percoll. The chloroplasts appear compact and highly refractile by phase contrast or Hoffmann contrast microscopy. Upon incubation with [3H]leucine or [35S]methionine, the chloroplasts incorporate the amino acids into protein at rates that are 100-fold faster than we had previously observed with Euglena and up to 8-fold faster than with chloroplasts of spinach. Euglena chloroplasts prepared by the current procedure are thus qualitatively superior to those previously available from Euglena and at least as active in protein synthesis as chloroplasts from higher plants.

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Selected References

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

  1. Bottomley W., Spencer D., Whitfeld P. R. Protein synthesis in isolated spinach chloroplasts: comparison of light-driven and ATP-driven synthesis. Arch Biochem Biophys. 1974 Sep;164(1):106–117. doi: 10.1016/0003-9861(74)90012-5. [DOI] [PubMed] [Google Scholar]
  2. Brown R. D., Haselkorn R. The isolation of Euglena gracilis chloroplasts uncontaminated by nuclear DNA. Biochim Biophys Acta. 1972 Jan 18;259(1):1–4. doi: 10.1016/0005-2787(72)90467-4. [DOI] [PubMed] [Google Scholar]
  3. Carell E. F. Studies on chloroplast development and replication in Euglena. I. Vitamin B12 and chloroplast replication. J Cell Biol. 1969 May;41(2):431–440. doi: 10.1083/jcb.41.2.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EISENSTADT J. M., BRAWERMAN G. THE PROTEIN-SYNTHESIZING SYSTEMS FROM THE CYTOPLASM AND THE CHLOROPLASTS OF EUGLENA GRACILIS. J Mol Biol. 1964 Dec;10:392–402. doi: 10.1016/s0022-2836(64)80060-7. [DOI] [PubMed] [Google Scholar]
  5. Freyssinet G., Harris G. C., Nasatir M., Schiff J. A. Events Surrounding the Early Development of Euglena Chloroplasts: 14. Biosynthesis of Cytochrome c-552 in Wild Type and Mutant Cells. Plant Physiol. 1979 May;63(5):908–915. doi: 10.1104/pp.63.5.908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Knight G. J., Price C. A. Measurements of s-p coordinates during the development of Euglena chloroplasts. Biochim Biophys Acta. 1968 May;158(2):283–285. doi: 10.1016/0304-4165(68)90142-6. [DOI] [PubMed] [Google Scholar]
  7. Morgenthaler J. J., Mendiola-Morgenthaler L. Synthesis of soluble, thylakoid, and envelope membrane proteins by spinach chloroplasts purified from gradients. Arch Biochem Biophys. 1976 Jan;172(1):51–58. doi: 10.1016/0003-9861(76)90046-1. [DOI] [PubMed] [Google Scholar]
  8. Morgenthaler J. J., Price C. A. Photosynthetic activity of spinach chloroplasts after isopycnic centrifugation in gradients of silica. Plant Physiol. 1974 Oct;54(4):532–534. doi: 10.1104/pp.54.4.532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Price C. A., Bourke M. E. "Spheroplasts" prepared from Euglena gracilis by proteolysis. J Protozool. 1966 Aug;13(3):474–477. doi: 10.1111/j.1550-7408.1966.tb01943.x. [DOI] [PubMed] [Google Scholar]
  10. Rawson J. R., Stutz E. Isolation and characterization of Euglena gracilis cytoplasmic and chloroplast ribosomes and their ribosomal RNA components. Biochim Biophys Acta. 1969 Oct 22;190(2):368–380. doi: 10.1016/0005-2787(69)90087-2. [DOI] [PubMed] [Google Scholar]
  11. Salisbury J. L., Vasconcelos A. C., Floyd G. L. Isolation of Intact Chloroplasts of Euglena gracilis by Isopycnic Sedimentation in Gradients of Silica. Plant Physiol. 1975 Sep;56(3):399–403. doi: 10.1104/pp.56.3.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Vasconcelos A. C. Synthesis of Proteins by Isolated Euglena gracilis Chloroplasts. Plant Physiol. 1976 Dec;58(6):719–721. doi: 10.1104/pp.58.6.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Vasconcelos A., Pollack M., Mendiola L. R., Hoffmann H. P., Brown D. H., Price C. A. Isolation of Intact Chloroplasts from Euglena gracilis by Zonal Centrifugation. Plant Physiol. 1971 Feb;47(2):217–221. doi: 10.1104/pp.47.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]

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