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Characterization of a light-dependent glutamate synthase activity in Chlamydomonas reinhardtii

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Abstract

Photosynthetically active vesicles prepared from Chlamydomonas reinhardtii retained a light-dependent glutamate synthase activity which was highly specific for 2-oxoglutarate (Km=2.1 mM) and L-glutamine (Km=0.9 mM) as amido group acceptor and donor respectively. This activity was inhibited by azaserine, p-hydroxymercuribenzoate and 3-(p-chlorophenyl)-1,1-dimethyl urea.

Light-dependent synthesis of glutamate was also obtained by coupling Chlamydomonas photosynthetic particles to purified ferredoxin-glutamate synthase, using ascorbate and 2,6-dichlorophenol-indophenol as electron donor. This system was also specific for 2-oxoglutarate (Km=1 mM) and L-glutamine (Km=0.8 mM) as substrates, and was stimulated by dithioerythritol. Azaserine and p-hydroxymercuribenzoate, but not 3-(p-chlorophenyl)-1,1-dimethyl urea, inhibited the reconstituted activity; high concentrations of 2-oxoglutarate were inhibitory.

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Abbreviations

A:

Absorbance

CCP:

p-Trichlorometoxi-carbonylcyanide-phenylhydrazone

Chl:

Chlorophyll

CMU:

3-(p-Chlorophenyl)-1,1-dimethyl urea

DPIP:

2,6-Dichlorophenol-indophenol

DTE:

Dithioerythritol

MSX:

L-Methionine, D-L, sulfoximine

MV:

Methyl viologen

References

  1. Anderson JW and Done J (1977) A polarographic study of glutamate synthase activity in isolated chloroplast. Plant Physiol 60: 354–359

    Google Scholar 

  2. Anderson JW and Walker DA (1983) Oxygen evolution by a reconstituted spinach chloroplast system in the presence of L-glutamine and 2-oxoglutarate. Planta 159: 77–83

    Google Scholar 

  3. Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenol oxidase in Beta vulgaris. Plant Physiol 24: 1–15

    Google Scholar 

  4. Dry IB and Wiskich JT (1983) Characterization of dicarboxylate stimulation of ammonia, glutamine and 2-oxoglutarate-dependent O2 evolution in isolated pea chloroplasts. Plant Physiol 72: 291–296.

    Google Scholar 

  5. Dry IB and Wiskich JT (1983) Measurement of in vitro glutamate synthase activity using an oxygen-evolving reconstituted chloroplast system. FEBS Lett 151: 31–35

    Google Scholar 

  6. Florencio F and Vega JM (1983) Separation, purification, and characterization of two isoforms of glutamine synthetase from Chlamydomonas reinhardii. Z Naturforsch 38c: 531–538

    Google Scholar 

  7. Galván F, Márquez AJ and Vega JM (1984) Purification and molecular properties of ferredoxin-glutamate synthase from Chlamydomonas reinhardii. Planta 162: 180–187

    Google Scholar 

  8. Galván F, Márquez AJ and Fernández E (1985) Physicochemical properties of ferredoxin from Chlamydomonas reinhardii. Z Naturforsch 40: 373–378

    Google Scholar 

  9. Wallsgrove RM, Keys AJ, Lea PJ and Miflin BJ (1983) Photosynthesis, photorespiration and nitrogen metabolism. Plant Cell Environment 6: 301–309

    Google Scholar 

  10. Márquez AJ (1985) Ph. D. Thesis, University of Seville, Spain

  11. Márquez AJ, Galván F and Vega JM (1984) Purification and characterization of NADH-glutamate synthase from Chlamydomonas reinhardii. Plant Sci Lett 34: 305–314

    Google Scholar 

  12. Márquez AJ, Galván F and Vega JM (1986) Utilization of ammonium by mutant and wild types Chlamydomonas reinhardii. Studies on the glutamate synthase activity. J Plant Physiol 124: 95–102

    Google Scholar 

  13. Márquez AJ, Gotor C, Romero LC, Galván F and Vega JM (1986) Ferredoxin-glutamate synthase from Chlamydomonas reinhardii. Prosthetic groups and preliminary studies of mechanism. Int J Biochem 18: 531–535

    Google Scholar 

  14. Matoh T, Ida S and Takahashi E (1980) A rapid and sensitive assay for ferredoxin-glutamate synthase. Bull Res Inst Food Sci, Kyoto Univ. 43: 1–6

    Google Scholar 

  15. McKenzie GH, Ch'ng AL and Gayler KR (1979) Glutamine synthetase/glutamine: α-ketoglutarate aminotransferase in chloroplast from the marine alga Caulerpa simpliciuscula. Plant Physiol 63: 578–582

    Google Scholar 

  16. Suzuki A, Jacquot JP, Martin F and Gadal P (1982) Light-dependent activity of glutamate synthase in vitro. Biochem. Biophys Res Commun 106: 65–71

    Google Scholar 

  17. Tischner R and Schmidt A (1982) A thioredoxin-mediated activation of glutamine synthetase and glutamate synthase in synchronous Chlorella sorokiniana. Plant Physiol 70: 113–116.

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Bioquímica, Facultad de Química, Apartado 553, 41080, Sevilla, Spain

    Antonio J. Márquez, M. Angeles Serra & José M. Vega

Authors
  1. Antonio J. Márquez
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  2. M. Angeles Serra
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  3. José M. Vega
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Márquez, A.J., Serra, M.A. & Vega, J.M. Characterization of a light-dependent glutamate synthase activity in Chlamydomonas reinhardtii . Photosynth Res 12, 73–81 (1987). https://doi.org/10.1007/BF00019152

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  • DOI: https://doi.org/10.1007/BF00019152

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