Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1967 Oct;42(10):1447–1456. doi: 10.1104/pp.42.10.1447

Photosynthetic Reactions by Lysed Protoplasts and Particle Preparations from the Blue-Green Alga, Phormidium luridum 1,2

John Biggins 1
PMCID: PMC1086745  PMID: 16656676

Abstract

Reactions of photosynthetic electron transport and photophosphorylation were studied in preparations from the blue-green alga, Phormidium luridum. Osmotic lysis of protoplasts proved to be a superior technique for the production of cell-free preparations with high enzymatic activity. Such lysed protoplasts sustain high rates of photophosphorylation coupled to the photo-reduction of NADP+ or ferricyanide. P/2e− ratios close to unity were routinely observed. The same preparations, and also those prepared by grinding the cells in solutions containing sucrose or ethylene glycol, are active in cyclic photophosphorylation mediated by phenazine methosulfate or dichloro-phenolindophenol. The particles prepared by grinding the cells are, however, inactive in non-cyclic photophosphorylation.

Extensive washing of the membranes with solutions containing sucrose removes the majority of the residual soluble fraction of the algal cell which includes cytochromes C554 and C549 and phycocyanin. Cyclic photophosphorylation activity is unimpaired by this treatment, but is abolished when the membranes are washed with very dilute buffers. This activity is restored by the addition of a soluble protein which is not a known redox constituent such as cytochrome C554 or plastocyanin, and may be a coupling factor.

Analysis of the well-washed membranes by low temperature (77°K) difference spectrophotometry reveals the presence of cytochrome b6 and a bound form of cytochrome C554 in proportions similar to that found in higher plant chloroplasts. The concentration of the membrane-bound cytochrome C554, relative to cytochrome b6 is not altered by extensive washing, sonication or treatment with 1% digitonin. This indicates that this cytochrome is an integral component of the cytoplasmic lamellae and we suggest that it is of functional significance. The soluble form of cytochrome C554, which is present in concentrations about 3-fold higher than the bound form, depending upon growth conditions, is not essential for cyclic photophosphorylation. The concentration of cytochrome b6: chlorophyll a was found to be 1:500.

Under the conditions employed, we were unable to detect a bound form of the low potential cytochrome C549.

Full text

PDF
1447

Selected References

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

  1. AVRON M. Photophosphorylation by swiss-chard chloroplasts. Biochim Biophys Acta. 1960 May 20;40:257–272. doi: 10.1016/0006-3002(60)91350-0. [DOI] [PubMed] [Google Scholar]
  2. AVRON M., SHAVIT N. A SENSITIVE AND SIMPLE METHOD FOR DETERMINATION OF FERROCYANIDE. Anal Biochem. 1963 Dec;6:549–554. doi: 10.1016/0003-2697(63)90149-0. [DOI] [PubMed] [Google Scholar]
  3. Biggins J., Park R. B. Physical Properties of Spinach Chloroplast Lamellar Proteins. Plant Physiol. 1965 Nov;40(6):1109–1115. doi: 10.1104/pp.40.6.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. ESTABROOK R. W. The low temperature spectra of hemoproteins. I. Apparatus and its application to a study of cytochrome c. J Biol Chem. 1956 Dec;223(2):781–794. [PubMed] [Google Scholar]
  5. FREDRICKS W. W., JAGENDORF A. T. A SOLUBLE COMPONENT OF THE HILL REACTION IN ANACYSTIS NIDULANS. Arch Biochem Biophys. 1964 Jan;104:39–49. doi: 10.1016/s0003-9861(64)80032-1. [DOI] [PubMed] [Google Scholar]
  6. Gerhardt B., Trebst A. Photosynthetische Reaktionen in lyophilisierten Zellen der Blaualge Anacystis. Z Naturforsch B. 1965 Sep;20(9):879–889. [PubMed] [Google Scholar]
  7. Gorman D. S., Levine R. P. Photosynthetic Electron Transport Chain of Chlamydomonas reinhardi VI. Electron Transport in Mutant Strains Lacking Either Cytochrome 553 or Plastocyanin. Plant Physiol. 1966 Dec;41(10):1648–1656. doi: 10.1104/pp.41.10.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorman D. S., Levine R. P. Photosynthetic Electron Transport Chain of Chlamydomonas reinhardi. IV. Purification and Properties of Plastocyanin. Plant Physiol. 1966 Dec;41(10):1637–1642. doi: 10.1104/pp.41.10.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorman D. S., Levine R. P. Photosynthetic Electron Transport Chain of Chlamydomonas reinhardi. V. Purification and Properties of Cytochrome 553 and Ferredoxin. Plant Physiol. 1966 Dec;41(10):1643–1647. doi: 10.1104/pp.41.10.1643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HILL R., BENDALL F. Crystallization of a photosynthetic reductase from a green plant. Nature. 1960 Jul 30;187:417–417. doi: 10.1038/187417a0. [DOI] [PubMed] [Google Scholar]
  11. HOLTON R. W., MYERS J. CYTOCHROMES OF A BLUE-GREEN ALGA: EXTRACTION OF A C-TYPE WITH A STRONGLY NEGATIVE REDOX POTENTIAL. Science. 1963 Oct 11;142(3589):234–235. doi: 10.1126/science.142.3589.234. [DOI] [PubMed] [Google Scholar]
  12. Holton R. W., Myers J. Water-soluble cytochromes from a blue-green alga. I. Extraction, purification, and spectral properties of cytochromes C (549, 552, and 554, Anacystis nidulans). Biochim Biophys Acta. 1967 Mar 8;131(2):362–374. doi: 10.1016/0005-2728(67)90150-8. [DOI] [PubMed] [Google Scholar]
  13. PERINI F., KAMEN M. D., SCHIFF J. A. IRON-CONTAINING PROTEINS IN EUGLENA. I. DETECTION AND CHARACTERIZATION. Biochim Biophys Acta. 1964 Jul 29;88:74–90. doi: 10.1016/0926-6577(64)90155-x. [DOI] [PubMed] [Google Scholar]
  14. SAN PIETRO A., LANG H. M. Photosynthetic pyridine nucleotide reductase. I. Partial purification and properties of the enzyme from spinach. J Biol Chem. 1958 Mar;231(1):211–229. [PubMed] [Google Scholar]
  15. SUSOR W. A., DUANE W. C., KROGMANN D. W. STUDIES ON PHOTOSYNTHESIS USING CELL FREE PREPARATIONS OF BLUE-GREEN ALGAE. Rec Chem Prog. 1964 Sep;25:197–208. [PubMed] [Google Scholar]
  16. SUSOR W. A., KROGMANN D. W. HILL ACTIVITY IN CELL-FREE PREPARATIONS OF A BLUE-GREEN ALGA. Biochim Biophys Acta. 1964 Jul 29;88:11–19. doi: 10.1016/0926-6577(64)90150-0. [DOI] [PubMed] [Google Scholar]
  17. Sauer K., Biggins J. Action spectra and quantum yields for nicotinamide--adenine dinucleotide phosphate reduction by chloroplasts. Biochim Biophys Acta. 1965 May 25;102(1):55–72. doi: 10.1016/0926-6585(65)90202-5. [DOI] [PubMed] [Google Scholar]
  18. TAGAWA K., TSUJIMOTO H. Y., ARNON D. I. Role of chloroplast ferredoxin in the energy conversion process of photosynthesis. Proc Natl Acad Sci U S A. 1963 Apr;49:567–572. doi: 10.1073/pnas.49.4.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wessels J. S. Isolation of a chloroplast fragment fraction with NADP+-photoreducing activity dependent on plastocyanin and independent of cytochrome f. Biochim Biophys Acta. 1966 Nov 8;126(3):581–583. doi: 10.1016/0926-6585(66)90016-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES