Abstract
In suspensions of the green alga Chlorella fusca the influence of high pH and high ethylene-diamine-tetraacetic acid concentrations in the external medium, of French-press and perchloric acid extraction of the cells and of alkalization of the intracellular pH on the polyphosphate signal in 31P-nuclear magnetic resonance (31P NMR) spectra was investigated.
The results show that part of the polyphosphates of asynchronous Chlorella cells are located outside the cytoplasmic membrane and complexed with divalent metal-ions. These polyphosphates are tightly bound to the cell wall and/or the cytoplasmic membrane and are not susceptible to hydrolyzation by strong acid at room temperature, in contrast to the intracytoplasmic polyphosphates.
Upon alkalization of the internal pH of Chlorella cells, polyphosphates, previously not visible in the spectra become detectable by 31P-NMR-spectroscopy. 31P-NMR spectroscopic monitoring of polyphosphates during gradual alkalization of the extra-and intracellular space is proposed as a quick method for the estimation of the cellular polyphosphate content and distribution.
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Abbreviations
- CCCP:
-
Carbonylcyanide-m-chlorophenyl-hydrazone
- NTP/NDP:
-
Nucleotide triphosphate/-diphosphate
- PCA:
-
Perchloric acid
- 31P-NMR:
-
31P-nuclear magnetic resonance
- PolyP:
-
polyphosphates
- PP1, PP2, PP3 :
-
terminal, second and third phosphate residue of polyphosphates, respectively
- PP4 :
-
core phosphate residues of polyphosphates
References
Atkinson AW, John PCL, Gunning BES (1974) The growth and division of the single mitochondrion and other organelles during the cell cycle of Chlorella, studied by quantitative stereology and three dimensional reconstruction. Protoplasma 81:77–109
Baker AL, Schmidt RR (1963) Intracellular distribution of phosphorus during synchronous growth of Chlorella pyrenoidosa. Biochim Biophys Acta 74:75–83
Baker AL, Schmidt RR (1964) Further studies on the intracellular distribution of phosphorus during synchronous growth of Chlorella pyrenoidosa. Biochim Biophys Acta 82:336–342
Den Hollander JA, Ugurbil K, Brown TR, Shulman RG (1981) Phosphorus-31 nuclear magnetic resonance studies of the effect of oxygen upon glycolysis in yeast. Biochemistry 20:5871–5880
Elgavish A, Elgavish GA, Halmann M, Berman T, Shomer I (1980) Intracellular phosphorus pools in intact algal cells. 31P-NMR and transmission electron microscopy studies. FEBS Lett 117:137–142
Florentz M, Granger P, Hartemann P (1984) Use of 31P nuclear magnetic resonance spectroscopy and electron microscopy to study phosphorus metabolism of microorganisms from wastewaters. Appl Environ Microbiol 47:519–525
Gadian DG (1982) Nuclear magnetic resonance and its applications to living systems. Oxford Univ Press. New York
Grimme LH, Boardman NK (1972) Photochemical activities of a particle fraction P1 obtained from the green alga Chlorella fusca. Biochem Biophys Res Commun 49:1617–1623
Gupta RK, Gupta P, Moore RD (1984) NMR studies of intracellular metal ions in intact cells and tissues. Ann Rev Biophys Bioeng 13:221–246
Kallas T, Dahlquist FW (1981) Phospohorus-31 nuclear magnetic resonance analysis of internal pH during photosynthesis in the cyanobacterium Synechococcus. Biochemistry 20:5900–5907
Kulaev IS (1979) The biochemistry of inorganic polyphosphates. Wiley and Sons, Chichester New York Brisbane Toronto
Kulaev IS, Vagaboy VM (1983) Polyphosphate metabolism in microorganisms. Adv Microb Physiol 24:83–171
Miyachi S, Miyachi S (1961) Modes of formation of phosphate compounds and their turnover in Chlorella cells during the process of life cycle as studied by the technique of synchronous culture. Plant Cell Physiol 2:415–424
Miyachi S, Tamiya H (1961) Distribution and turnover of phosphate compounds in growing Chlorella cells. Plant Cell Physiol 2:405–414
Miyachi S, Kanai R, Mihara S, Miyachi S, Aoki S (1964) Metabolic roles of inorganic polyphosphates in Chlorella cells. Biochim Biophys Acta 93:625–634
Navon G, Shulman RG, Yamana T, Eccleshall TR, Lam KB, Baronofsky JJ, Marmur J (1979) Phosphorus-31 nuclear magnetic resonance studies of wild-type and glycolytic pathway mutants of Saccharomyces cerevisiae. Biochemistry 18:4487–4499
Nicolay K, Scheffers WA, Bruinenberg PM, Kaptein R (1982) Phosphorus-31 nuclear magnetic resonance studies of intracellular pH, phosphate compartmentation and phosphate transport in yeasts. Arch Microbiol 133:83–89
Nicolay K, Scheffers WA, Bruinenberg PM, Kaptein R (1983) In vivo 31P-NMR studies on the role of the vacuole in phosphate metabolism in yeasts. Arch Microbiol 134:270–275
Ostrovskii DN, Sepetov NF, Reshetnyak VI, Siberl'dina LA (1980) Investigations of the localization of polyphosphates in cells of microorganisms by the method of high resolution 31P-NMR-145.78 MHz. Biochemistry USSR 45:517–525
Peverly JH, Adamec J, Parthasarathy MV (1978) Association of potassium and some other monovalent cations with occurrence of polyphosphate bodies in Chlorella pyrenoidosa. Plant Physiol 62:120–126
Sianoudis J, Mayer A, Leibfritz D (1984) Investigation of intracellular phosphate pools of the green alga Chlorella using 31P-nuclear magnetic resonance. Org Magn Resonance 22:364–368
Sianoudis J, Naujokat T, Mayer A, Grimme LH, Leibfritz D (1985) 31P-NMR studies of the phosphate metabolism of the green alga Chlorella fusca. Govil G, Khetrapal CL and Saran A (eds) Magnetic resonance in biology and medicine. Tata McGraw-Hill Publishing Company Limited, New Delhi, India, pp 429–438
Sianoudis J, Küsel AC, Naujokat T, Offermann W, Mayer A, Grimme LH, Leibfritz D (1985a) Respirational activity of Chlorella fusca monitored by in vivo P-31 NMR. Eur Biophys J (in press)
Tantawy MM, Grimme LH (1982) The bleaching induced by the phenylpyridazinone herbicide metflurazon in Chlorella is a metabolic and reversible process. Pest Biochem Physiol 18:304–314
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Sianoudis, J., Küsel, A.C., Mayer, A. et al. Distribution of polyphosphates in cell-compartments of Chlorella fusca as measured by 31P-NMR-spectroscopy. Arch. Microbiol. 144, 48–54 (1986). https://doi.org/10.1007/BF00454955
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DOI: https://doi.org/10.1007/BF00454955