Abstract
Intracellular ion concentration and respiratory activity in the marine cyanobacterium Spirulina subsalsa was analyzed during cell transition from saline to hypersaline medium. During salt upshock, an early phase of Na+ and Cl- influx was observed, followed by an adaptation phase where both Na+ and Cl- were excluded from the cell. Respiration in intact cells was enhanced during salt upshock. S. subsalsa spheroplasts exhibited a high rate of O2 uptake, which was further enhanced in cells grown in hypersaline medium, upon addition of NaCl to the assay mixture. This effect was found to be specific to sodium ions. Plasma membrane fractions from cells grown in hypersaline medium exhibited a high rate of cytochrome oxidase activity, which was further stimulated by NaCl, and was sensitive to DCCD. Immunoblot analysis of Spirulina plasma membrane polypeptides with anti-cytochrome oxidase serum demonstrated high content of 53.4 kDa polypeptide of cytochrome oxidase, which was enriched in membranes obtained from hypersaline Spirulina cells. The enhanced respiration, and more specifically the enrichment of cytochrome oxidase activity in salt-adapted cells in situ, as well as its stimulation by NaCl in vitro and inhibition by DCCD, suggest that cytochrome oxidase is involved in the extrusion of sodium ions from cells of the salt-tolerant Spirulina subsalsa.
Similar content being viewed by others
Abbreviations
- DCCD:
-
dicyclohexylcarbodiimide
- CCCP:
-
carbonylcyanide m-chlorophenyl hydrazone
- TMPD:
-
N, N, N′, N′, tetramethyl p-phenylenediamine dichloride
References
Abeliovich A, Gan J (1982) Factors affecting motility of Spirulina subsalsa. In: Papageorgiou GC, Packer L (eds) Photosynthetic prokaryotes: cell differentiation and function. Elsevier, New York, pp 353–362
Babcock GT, Callahan PM (1983) Redox-linked hydrogen bond strength changes in cytochrome a: implications for a cytochrome oxidase proton pump. Biochemistry 22: 2314–2319
Blumwald E, Wolosin JM, Packer L (1984) Na+/H+ Exchange in the cyanobacterium Synechocossus 6311. Biochem Biophys Res Commun 122: 452–459
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72: 248–254
Burnette WN (1981) “Western Blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody radiodinated protein A. Anal Biochem 112: 195–203
Casey RP, Thelen M; Azzi A (1980) Dicyclohexylcarbodiimide binds specifically and covalently to cytochrome c oxidase while inhibiting H+-translocating activity. J Biol Chem 25: 3994–4000
Delaney SF (1984) Spheroplasts of Synechococcus PCC 6301. J Gen Microbiol 130: 2771–2773
Erber WWA, Nitschmann WH, Muchl R, Peschek GA (1986) Endogenous energy supply to the plasma membrane of dark aerobic cyanobacterium Anacystis nidulans: ATPase-independent efflux of H+ and Na+ from respiring cells. Arch Biochem Biophys 247: 28–39
Finel M (1989) Oligomeric structure and subunit requirement of proton-translocating cytochrome oxidase. PH D Thesis, University of Helsinki, Faculty of 111
Fry IV, Huflejt M, Erber WWA, Peschek GA, Packer L (1986) The role of respiration during adaptation of the freshwater cyanobacterium Synechoccus 6311 to salinity. Arch Biochem Biophys 244: 686–691
Gabbay R, Tel-Or E (1985) Cyanobacterial biomass production in saline media. Plant Soil 89: 107–116
Gabbay-Azaria R, Tel-Or E, Schonfeld M (1988) Glycinebetaine as an osmoregulant and osmoprotectant in the marine cyanobacterium Spirulina subsalsa. Arch Biochem Biophys 261: 333–339
Kienzel PF, Peschek GA (1982) Oxidation of c-type cytochromes by the membrane-bound cytochrome oxidase (cytochrome aa3) of blue-green algae. Plant Physiol 69: 580–584
Kirst O (1989) Salinity tolerance of eukaryotic marine algae. Ann Rev Plant Physiol Plant Mol Biol 40: 21–53
Krab K, Wikstrom M (1987) Principles of coupling between electron transfer and proton translocation with special reference to proton translocation mechanisms in cytochrome oxidase. Biochim Biophys Acta 895: 25–39
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Lauraeus M, Haltia T, Saraste M, Wikstrom M (1991) Bacillus subtilis expresses two kinds of haem-A-containing terminal oxidases. Eur J Biochem 197: 699–705
MacKinney G (1941) Absorption of light by chlorophyll solutions. J Biol Chem 140: 315–322
Markwell MAK, Haas MS, Bieber LL, Tolbert NE (1978) A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87: 206–210
Molitor V, Peschek GA (1986) Respiratory electron transport in plasma and thylakoid membrane preparations from the cyanobacterium Anacystis nidulans. FEBS Lett 195: 145–150
Molitor V, Walter E, Peschek GA (1986) Increased levels of cytochrome oxidase and sodium-proton antiporter in the plasma membrane of Anacystis nidulans after growth in sodium-enriched media. FEBS Lett 204: 251–256
Nitschmann WH, Peschek GA (1985) Modes of proton translocation across the cell membrane of respiring cyanobacteria. Arch Microbiol 141: 330–336
Nitschmann WH, Schmetterer G, Muchl R, Peschek GA (1982) Active sodium extrusion reduces net efficiencies of oxidative phosphorylation in the strictly photoautotrophic cyanobacterium Anacystis nidulans. Biochim Biophys Acta 682: 293–296
Omata T, Murata N (1985) Electron transport reactions in cytoplasmic and thylakoid membranes prepared from the cyanobacterium (blue-green algae) Anacystis nidulans and Synechosystic PCC 6714. Biochim Biophys Acta 810: 354–361
Paschinger H (1977) Sodium chloride stimulated respiration of Anacystis nidulans. Z Allg Mikrobiol 17: 377–379
Packer L, Spath S, Martin JB, Roby C, Bligny R (1987) 23Na and 31P NMR studies of the effects of salt stress on the freshwater cyanobacterium Synechococcus 6311. Arch Biochim Biophys 256: 354–361
Peschek GA (1981 a) Occurrence of cytochrome aa3 in Anacystis nidulans. Biochim Biophys Acta 635: 470–475
Peschek GA (1981 b) Spectral properties of a cyanobacterial cytochrome c oxidase: evidence for cytochrome aa3. Biochem Biophys Res Commun 98: 72–79
Peschek GA (1983) Proton pum coupled to cytochrome c oxidase in the cyanobacterium Anacystis nidulans. J Bacteriol 153: 539–542
Peschek GA (1984) Characterization of the proton translocating cytochrome c oxidase activity in the plasma membrane of intact Anacystic nidulans spheroplasts. Plant Physiol 75: 968–973
Peschek GA (1987) Respiratory electron transport. In: Fay P, VanBaalen C (eds) The cyanobacteria. Elsevier, New York, pp 119–161
Peschek GA, Molitor V, Trnka M, Waystyn M, Erber W (1988a) Characterization of the cytochrome c oxidase in isolated and purified plasma and thylakoid membranes from cyanobacteria. Methods Enzymol 167: 437–449
Peschek A, Nitschmann WH, Czerny T (1988b) Respiratory proton extrusion and plasma membrane energization. Methods Enzymol 167: 361–379
Peschek GA, Schmetterer G, Lauritsch G, Muchl R, Kinzl PF, Nitschmann WH (1982) Proton-pumping cytochrome oxidase in the cytoplasmic membrane of Anacystis nidulans. In: Papageorgio CG, Packer L (eds) Photosynthetic prokaryotes: cell differentiation and function. Elsevier, New York, pp. 147–162
Pool RK (1988) Bacterial cytochrome oxidases. In: Christopher A (ed) Bacterial energy transduction. Academic Press, London, pp 231–291
Prochaska L, Bisson R, Capaldi RA, Steffens GCM, Buse G (1981) Inhibition of cytochrome c oxidase function by dicyclohexylcarbodiimide. Biochim Biophys Acta 637: 360–373
Prochaska LJ, Fink PS (1987) On the role of subunit III in proton translocation in cytochrome c oxidase. J Bioenerg Biomebr 19: 143–166
Reynafarje B, Costa LE, Lehninger AL (1986) Upper and lower limits of the proton stoichiometry of cytochrome c oxidation in rat liver mitoplasts. J Biol Chem 261: 8254–8262
Steinrucke P, Steffens GCM, Panskus G, Buse G, Ludwig B (1987) Subunit II of cytochrome oxidase from Paracoccus denitrificans. DNA sequence, gene expression and the protein. Eur J Biochem 167: 431–439
Stone BB, Boussiba S, Gibson J (1988) Penicillin-binding proteins in the unicellular cyanobacterium Synechococcus sp. strain R2 (PCC 1942): implications for membrane identification. J Gen Microbiol 134: 2951–2958
Tel-Or E (1979) Adaptation to salt of the photosynthetic apparatus in cyanobacteria. FEBS Lett 110: 253–256
Trnka M, Peschek GA (1986) Immunological identification of aa3-type cytochrome oxidase in membrane preparations of the cyanobacterium Anacystis nidulans. Biochem Biophys Res Commun 136: 235–241
Vonshak A, Guy R, Guy M (1988) The response of filamentous cyanobacterium Spirulina platensis to salt stress. Arch Microbiol 150: 417–420
Wastyn M, Achatz A, Molitor V, Peschek GA (1988) Respiratory activities and aa3-type cytochrome oxidase in plasma and thylakoid membranes from vegetative cells and heterocysts of the cyanobacterium Anabaena ATCC 29413. Biochim Biophys Acta 935: 217–224
West IC, Mitchell R, Moody AJ, Mitchell P (1987) Proton translocation by cytochrome oxidase in (antimycin + myxothiazol)-treated rat liver mitochondria using ferrocyanide or hexammineruthenium as electron donor. FEBS Lett 222: 235–245
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gabbay-Azaria, R., Schonfeld, M., Tel-Or, S. et al. Respiratory activity in the marine cyanobacterium Spirulina subsalsa and its role in salt tolerance. Arch. Microbiol. 157, 183–190 (1992). https://doi.org/10.1007/BF00245288
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00245288