Summary
Photoautotrophically growing cultures of the fresh water cyanobacteriumAnacystis nidulans adapted to the presence of 0.4–0.5 M NaCl (about sea water level) with a lag phase of two days after which time the growth rate reassumed 80–90% of the control. Plasma and thylakoid membranes were separated from cell-free extracts of French pressure cell treatedAnacystis nidulans by discontinuous sucrose density gradient centrifugation and purified by repeated recentrifugation on fresh gradients. Identity of the plasma and thylakoid membrane fractions was confirmed by labeling of intact cells with impermeant protein markers prior to breakage and membrane isolation. Electron microscopy revealed that each type of membrane was obtained in the form of closed and perfectly spherical vesicles. Major changes in structure and function of the plasma membranes (and, to a much lesser extent, of the thylakoid membranes) were found to accompany the adaptation process. On the average, diameters of plasma membrane vesicles from salt adapted cells were only one-third of the diameters of corresponding vesicles from control cells. By contrast, the diameters of thylakoid membrane vesicles were the same in both cases.
Freeze-etching the cells and counting the number of membrane-intercalating particles on both protoplasmic and exoplasmic fracture faces of plasma and thylakoid membranes indicated a roughly 50% increase of the particle density in plasma membranes during the adaptation process while that in thylakoid membranes was unaffected. Comparison between particle densities on isolated membranes and those on corresponding whole cell membranes permitted an estimate as to the percentage of inside-out and right-side-out vesicles. Stereometric measurement of particle sizes suggested that two distinct sub-populations of the particles in the plasma membranes increased during the adaptation process, tentatively correlated to the cytochrome oxidase and sodium-proton antiporter, respectively. The effects of salt adaptation described in this paper were fully reversed upon withdrawal of the additional NaCl from the growth medium (“deadaptation”). Moreover, they were not observed when the NaCl was replaced by KCl.
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Abbreviations
- CM:
-
cytoplasmic or plasma membrane
- ICM:
-
intracytoplasmic or thylakoid membrane
- EF:
-
exoplasmic fracture face
- PF:
-
protoplasmic fracture face
- DABS:
-
diazobenzosulfonate; Hepes N-2-hydroxyethylpiperazine-N′-2-ethane-sulfonate
- PMSF:
-
phenylmethylsulfonylfluoride
References
Armond PA, Staehelin LA (1979) Lateral and vertical displacement of integral membrane proteins during lipid phase transition inAnacystis nidulans. Proc Natl Acad Sci USA 76: 1901–1905
Azaria-Gabbay R, Tel-Or E, Schonfeld M (1988) Mechanisms of salt tolerance in the marine cyanobacteriumSpirulina subsalsa. In: Mur LR, Burger-Wiersma T (eds) Abstracts 6th International Symposium on Phototrophic Prokaryotes, Noordwijkerhout, The Netherlands, p 155
Berg HC (1969) Sulfanilic acid diazonium salt: a label for the outside of the human erythrocyte membrane. Biochim Biophys Acta 183: 65–78
Blumwald E, Tel-Or E (1982 b) Structural aspects of theNostoc muscorum to salt. Arch Microbiol 132: 162–167
— — (1982 a) Osmoregulation and cell composition in salt adaptation ofNostoc muscorum. Arch Microbiol 132: 168–172
—, Mehlhorn JR, Packer L (1983 a) Ionic osmoregulation during salt adaptation of the cyanobacteriumSynechococcus 6311. Plant Physiol 73: 377–380
— — — (1983 b) Studies of osmoregulation in salt adaptation of cyanobacteria with ESR spin probe techniques. Proc Natl Apad Sci USA 80: 2599–2602
—, Wolosin JM, Packer L (1984) Na+/H+ exchange in the cyanobacteriumSynechococcus 6311. Biochem Biophys Res Commun 122: 452–459
Brethes D, Dulon D, Johannin G, Arrio B, Gulik-Krzywicki T, Chevalier J (1986) Study of electrokinetic properties of reconstituted sarcoplasmic reticulum vesicles. Arch Biochem Biophys 246: 355–365
Erber WWA, Nitschmann WH, Muchl R, Peschek GA (1986) Endogenous energy supply to the plasma membrane of dark aerobic cyanobacteriumAnacystis nidulans: ATPase-independent efflux of H+ and Na+ from respiring cells. Arch Biochem Biophys 247: 28–38
Fry IV, Huflejt M, Erber WWA, Peschek GA, Packer L (1986) The role of respiration during salt adaptation of the fresh water cyanobacteriumSynechococcus 6311. Arch Biochem Biophys 244: 686–691
Kratz WA, Myers J (1955) Nutrition and growth of several blue-green algae. Amer J Bot 42: 282–287
Lefort-Tran M, Pouphile M, Spath S, Packer L (1988) Cytoplasmic membrane changes during adaptation of the fresh water cyanobacteriumSynechococcus 6311 to salinity. Plant Physiol 87: 767–775
Molitor V, Peschek GA (1986) Respiratory electron transport in plasma and thylakoid membrane preparations from the cyanobacteriumAnacystis nidulans. FEBS Lett 195: 145–150
—, Erber WWA, Peschek GA (1986) Increased levels of cytochrome oxidase and sodium-proton antiporter in the plasma membrane ofAnacystis nidulans after growth in sodium-enriched media. FEBS Lett 204: 251–256
—, Trnka M, Peschek GA (1987) Isolated and purified plasma and thylakoid membranes of the cyanobacteriumAnacystis nidulans contain immunologically cross-reactive aa3-type cytochrome oxidase. Curr Microbiol 14: 263–268
Muchl R, Peschek GA (1984) Valinomycin pulse induced phosphorylation of ADP in dark anaerobic cells of the cyanobacteriumAnacystis nidulans. Curr Microbiol 11: 179–182
Murata N, Omata T (1988) Isolation of cyanobacterial plasma membranes. Methods Enzymol 167: 245–251
Nitschmann WH, Peschek GA (1985) Modes of proton translocation across the cell membrane of respiring cyanobacteria. Arch Microbiol 141: 330–336
Peschek GA (1983) Proton pump coupled to cytochromec oxidase in the cyanobacteriumAnacystis nidulans. J Bacteriol 153: 539–542
— (1984) Characterization of the proton-translocating cytochromec oxidase in the plasma membrane of intactAnacystis nidulans spheroplasts. Plant Physiol 75: 278–284
—, Czerny T, Schmetterer G, Nitschmann WH (1985) Transmembrane proton electrochemical gradients in dark aerobic and anaerobic cells of the cyanobacterium (blue-green alga)Anacystis nidulans. Plant Physiol 79: 278–284
—, Molitor V, Trnka M, Wastyn M, Erber WWA (1988 a) Characterization of cytochromec oxidase in isolated and purified plasma and thylakoid membranes from cyanobacteria. Methods Enzymol 167: 437–449
—, Nitschmann WH, Czerny T (1988 b) Respiratory proton extrusion and plasma membrane energization. Methods Enzymol 167: 361–379
—, Hinterstoisser B, Wastyn M, Kuntner O, Pineau B, Missbichler A, Lang J (1989) Chlorophyll precursors in the plasma membrane of a cyanobacterium,Anacystis nidulans. J Biol Chem 264: 11827–11832
Reed RH, Richardson DL, Stewart WDP (1985 b) Na+ uptake and extrusion in the cyanobacteriumSynechocystis PCC 6714 in response to hypersaline treatment. Evidence for transient changes in plasmalemma Na+ permeability. Biochim Biophys Acta 814: 347–355
—, Warr SRC, Richardson DL, Moore DJ, Stewart WDP (1985 a) Multiphasic osmotic adjustment in a euryhaline cyanobacterium. FEMS Microbiol Lett 28: 225–229
—, Borowitzka LJ, Mackay MA, Chudek JA, Foster R, Warr SRC, Moore DJ, Stewart WDP (1986) Organic solute accumulation in osmotically stressed cyanobacteria. FEMS Microbiol Rev 39: 51–56
—, Richardson DL, Warr SRC, Stewart WDP (1984) Carbohydrate accumulation and osmotic stress in cyanobacteria. J Gen Microbiol 130: 1–4
Schmetterer G, Peschek GA, Sleytr UB (1983) Thylakoid degradation during photooxidative bleaching of the cyanobacteriumAnacystis nidulans. Protoplasma 115: 202–207
Tinberg HM, Melnick RL, Maguire J, Packer L (1974) Studies on mitochondrial proteins. II. Localization of components in the inner membrane: labeling with diazobenzenesulfonate, a non-penetrating probe. Biochim Biophys Acta 345: 118–128
Udenfriend S, Stein S, Böhlein P, Dairman W, Leimgruber W, Weigele M (1972) Fluorescamine: a reagent for assay of amino acids, peptides, proteins and primary amines in the picomole range. Science 178: 871–872
Verkleij AJ (1984) Lipid intramembranous particles. Biochim Biophys Acta 779: 43–63
Verwer W, Ververgaert PHJT, Leunissen-Bijvelt J, Verkleij AJ (1978) Particle aggregation in photosynthetic membranes of the blue-green algaAnacystis nidulans. Biochim Biophys Acta 504: 231–234
Vonshak A, Richmond A (1981) Photosynthetic and respiratory activity inAnacystis nidulans adapted to osmotic shock. Plant Physiol 68: 504–507
Wada H, Hirasawa R, Omata T, Murata N (1984) The lipid phase of thylakoid and cytoplasmic membranes from the blue-green algae (cyanobacteria)Anacystis nidulans andAnabaena variabilis. Pl Cell Physiol 25: 907–911
Wastyn M, Achatz A, Trnka M, Peschek GA (1987) Immunological and spectral characterization of partly purified cytochrome oxidase from the cyanobacteriumSynechocystis 6714. Biochem Biophys Res Commun 149: 102–111
—, Achatz A, Molitor V, Peschek GA (1988 a) Respiratory activities and aa3-type cytochrome oxidase in plasma and thylakoid membranes from vegetative cells and heterocysts of the cyanobacteriumAnabaena ATCC 29413. Biochim Biophys Acta 935: 217–224
—, Peschek GA (1988 b) Aspects of sidedness and regulation of cytochrome oxidase (aa3-type) in cyanobacterial membrane vesicles. EBEC Rep 5: 94
Weigele M, de Bernardo SL, Tengi JP, Leimgruber W (1972) A novel reagent for the fluorimetric assay of primary amines. J Am Chem Soc 94: 5927–5928
Yonetani T (1965) Studies on cytochrome oxidase III. Improved preparation and some properties. J Biol Chem 236: 1680–1688
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Dedicated to the memory of Professor Oswald Kiermayer
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Molitor, V., Kuntner, O., Sleytr, U.B. et al. The ultrastructure of plasma and thylakoid membrane vesicles from the fresh water cyanobacteriumAnacystis nidulans adapted to salinity. Protoplasma 157, 112–119 (1990). https://doi.org/10.1007/BF01322643
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DOI: https://doi.org/10.1007/BF01322643