Molecular and regulatory properties of glutamine synthetase from the phototrophic bacterium Rhodopseudomonas capsulata E1F1
10.1128/jb.162.2.804-809.1985
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The glutamine synthetase of the phototrophic bacterium Rhodopseudomonas capsulata E1F1 was purified to homogeneity by a procedure which used a single affinity chromatography step. Like enzymes from other photosynthetic procaryotes, native glutamine synthetase from R. capsulata E1F1 was found to be a dodecameric protein of approximately 660 kilodaltons with identical subunits of about 55 kilodaltons each. The Stokes radius and S20,w of the native enzyme were 8.35 nm and 19.20, respectively. The enzyme exhibited different aggregation states with detectable oligomers of 1, 2, 3, 4, 6, 8, 10, and 12 subunits. Disaggregation of the glutamine synthetase occurred after the native protein was subjected to electrophoresis in polyacrylamide gels, as well as occurring spontaneously at low ionic strength. Glutamine synthetase from R. capsulata E1F1 was regulated by an adenylylation-deadenylylation mechanism, and the adenylylation state of the protein depended on the nitrogen source, growth phase, and light intensity. Ammonia repressed glutamine synthetase, whereas glycine, serine, alanine, valine, and aspartate were noncompetitive inhibitors of the glutamine synthetase biosynthetic activity.

Bibliography

Caballero, F. J., Cejudo, F. J., Florencio, F. J., Cárdenas, J., & Castillo, F. (1985). Molecular and regulatory properties of glutamine synthetase from the phototrophic bacterium Rhodopseudomonas capsulata E1F1. Journal of Bacteriology, 162(2), 804–809.

Authors 5
  1. F J Caballero (first)
  2. F J Cejudo (additional)
  3. F J Florencio (additional)
  4. J Cárdenas (additional)
  5. F Castillo (additional)
References 40 Referenced 25
  1. Alef , K. , H. -J. Burkardt , H. -J. Horstmann , and W. G. Zumft . 1981 . Molecular characterization of glutamine synthetase from the nitrogen-fixing phototrophic bacterium Rhodopseudomonas palustris . Z. Naturforsch. 36 : 246 - 254 . (10.1515/znc-1981-3-411) / Z. Naturforsch. / Molecular characterization of glutamine synthetase from the nitrogen-fixing phototrophic bacterium Rhodopseudomonas palustris by Alef K. (1981)
  2. Alef , K. , and W. G. Zumft . 1981 . Regulatory properties of glutamine synthetase from the nitrogen-fixing phototrophic bacterium Rhodopseudomonas palustris . Z. Naturforsch. 36 : 784 - 789 . (10.1515/znc-1981-9-1015) / Z. Naturforsch. / Regulatory properties of glutamine synthetase from the nitrogen-fixing phototrophic bacterium Rhodopseudomonas palustris by Alef K. (1981)
  3. Bradford , M. M. 1976 . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding . Anal. Biochem. 72 : 248 - 254 . (10.1016/0003-2697(76)90527-3) / Anal. Biochem. / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding by Bradford M. M. (1976)
  4. Brewer J. M. A. J. Pesce and T. E. Spencer. 1974. Ultracentrifugation p. 161-215. In J. M. Brewer A. J. Pesce and R. B. Ashworth (ed.) Experimental techniques in biochemistry. Prentice-Hall Inc. Englewood Cliffs N.J.
  5. Davies , W. , and J. G. Ormerod . 1982 . Glutamine synthetase in Chlorobium limicola and Rhodospirillum rubrum . FEMS Microbiol. Lett. 13 : 75 - 78 . (10.1111/j.1574-6968.1982.tb08230.x) / FEMS Microbiol. Lett. / Glutamine synthetase in Chlorobium limicola and Rhodospirillum rubrum by Davies W. (1982)
  6. Deuel , T. F. , and S. Prusiner . 1974 . Regulation of glutamine synthetase from Bacillus subtilis by divalent cations, feedback inhibitors and L-glutamine . J. Biol. Chem. 249 : 257 - 264 . (10.1016/S0021-9258(19)43119-0) / J. Biol. Chem. / Regulation of glutamine synthetase from Bacillus subtilis by divalent cations, feedback inhibitors and L-glutamine by Deuel T. F. (1974)
  7. Dharmawerdene , M. W. N. , A. Haystead , and W. D. P. Stewart . 1973 . Glutamine synthetase of the nitrogen-fixing alga Anabaena cylindrica . Arch. Mikrobiol. 90 : 281 - 295 . (10.1007/BF00408924) / Arch. Mikrobiol. / Glutamine synthetase of the nitrogen-fixing alga Anabaena cylindrica by Dharmawerdene M. W. N. (1973)
  8. Engelhardt , H , and J. -H. Klemme . 1982 . Purification and structural properties of adenylylated and deadenylylated glutamine synthetase from Rhodopseudomonas sphaeroides . Arch. Microbiol. 133 : 202 - 205 . (10.1007/BF00415001) / Arch. Microbiol. / Purification and structural properties of adenylylated and deadenylylated glutamine synthetase from Rhodopseudomonas sphaeroides by Engelhardt H (1982)
  9. Florencio , F. J. , and J. M. Vega . 1983 . Separation, purification, and characterization of two isoforms of glutamine synthetase from Chlamydomonas reinhardii . Z. Naturforsch. 38 : 531 - 538 . (10.1515/znc-1983-7-806) / Z. Naturforsch. / Separation, purification, and characterization of two isoforms of glutamine synthetase from Chlamydomonas reinhardii by Florencio F. J. (1983)
  10. Fuchs , R. L. , and D. L. Keister . 1980 . Identification of two glutamine synthetases in Agrobacterium . J. Bacteriol. 141 : 996 - 998 . (10.1128/jb.141.2.996-998.1980) / J. Bacteriol. / Identification of two glutamine synthetases in Agrobacterium by Fuchs R. L. (1980)
  11. Hedrick , J. L. , and D. J. Smith . 1968 . Size and charge isomer separation and estimation of molecular weights of protein by disc gel electrophoresis . Arch. Biochem. Biophys. 126 : 155 - 164 . (10.1016/0003-9861(68)90569-9) / Arch. Biochem. Biophys. / Size and charge isomer separation and estimation of molecular weights of protein by disc gel electrophoresis by Hedrick J. L. (1968)
  12. Hillmer , P. , and K. Fahlbusch . 1979 . Evidence for an involvement of glutamine synthetase in regulation of nitrogenase activity in Rhodopseudomonas capsulata . Arch. Microbiol. 122 : 213 - 218 . (10.1007/BF00411363) / Arch. Microbiol. / Evidence for an involvement of glutamine synthetase in regulation of nitrogenase activity in Rhodopseudomonas capsulata by Hillmer P. (1979)
  13. Johansson , B. C. , and H. Gest . 1977 . Adenylylation/deadenylylation control of glutamine synthetase of Rhodopseudomonas capsulata . Eur. J. Biochem. 81 : 365 - 371 . (10.1111/j.1432-1033.1977.tb11960.x) / Eur. J. Biochem. / Adenylylation/deadenylylation control of glutamine synthetase of Rhodopseudomonas capsulata by Johansson B. C. (1977)
  14. Jovin , T. , A. Chrambach , and M. A. Naughton . 1964 . Apparatus for preparative temperature regulated polyacrylamide gel electrophoresis . Anal. Biochem. 9 : 351 - 364 . (10.1016/0003-2697(64)90192-7) / Anal. Biochem. / Apparatus for preparative temperature regulated polyacrylamide gel electrophoresis by Jovin T. (1964)
  15. Kaulen , H. , and J. -H. Klemme . 1983 . No evidence of covalent modification of glutamine synthetase in the thermophilic phototrophic bacterium Chloroflexus aurantiacus . FEMS Microbiol. Lett. 20 : 75 - 79 . (10.1111/j.1574-6968.1983.tb00092.x) / FEMS Microbiol. Lett. / No evidence of covalent modification of glutamine synthetase in the thermophilic phototrophic bacterium Chloroflexus aurantiacus by Kaulen H. (1983)
  16. Khanna , S. , and D. J. D. Nicholas . 1983 . Some properties of glutamine synthetase and glutamate synthase from Chlorobium vibrioforme f. thiosulfatophilum . Arch. Microbiol. 134 : 98 - 103 . (10.1007/BF00407939) / Arch. Microbiol. / Some properties of glutamine synthetase and glutamate synthase from Chlorobium vibrioforme f. thiosulfatophilum by Khanna S. (1983)
  17. Khanna , S. , and D. J. D. Nicholas . 1983 . Adenylylation of glutamine synthetase in Chlorobium vibrioforme f. thiosulfatophilum . FEMS Microbiol. Lett. 18 : 173 - 176 . (10.1111/j.1574-6968.1983.tb00473.x) / FEMS Microbiol. Lett. / Adenylylation of glutamine synthetase in Chlorobium vibrioforme f. thiosulfatophilum by Khanna S. (1983)
  18. Kingdon , H. S. , B. M. Shapiro , and E. R. Stadtman . 1967 . Regulation of glutamine synthetase. VIII. ATP:glutamine synthetase adenylyltransferase, an enzyme that catalyzes alterations in the regulatory properties of glutamine synthetase . Proc. Natl. Acad. Sci. U.S.A. 58 : 1703 - 1710 . (10.1073/pnas.58.4.1703) / Proc. Natl. Acad. Sci. U.S.A. / Regulation of glutamine synthetase. VIII. ATP:glutamine synthetase adenylyltransferase, an enzyme that catalyzes alterations in the regulatory properties of glutamine synthetase by Kingdon H. S. (1967)
  19. Lowry , 0. H. , M. J. Rosebrough , A. L. Farr , and R. J. Randall . 1951 . Protein measurement with the Folin phenol reagent . J. Biol. Chem. 193 : 265 - 275 . (10.1016/S0021-9258(19)52451-6) / J. Biol. Chem. / Protein measurement with the Folin phenol reagent by Lowry 0. (1951)
  20. Magasanik B. and D. M. Rothstein. 1980. The role of glutamine synthetase in the regulation of bacterial nitrogen metabolism p. 61-68. In J. Mora and R. Palacios (ed.) Glutamine: metabolism enzymology and regulation. Academic Press Inc. New York. (10.1016/B978-0-12-506040-0.50008-X)
  21. Martin , R. G. , and B. N. Ames . 1961 . A method of determining the sedimentation behavior of enzymes: application to protein mixtures . J. Biol. Chem. 236 : 1372 - 1379 . (10.1016/S0021-9258(18)64180-8) / J. Biol. Chem. / A method of determining the sedimentation behavior of enzymes: application to protein mixtures by Martin R. G. (1961)
  22. Moreno-Vivian , C. , F. J. Cejudo , J. Cardenas , and F. Castillo . 1983 . Ammonia assimilation pathways in Rhodopseudomonas capsulata ElFl . Arch. Microbiol. 136 : 147 - 151 . (10.1007/BF00404790) / Arch. Microbiol. / Ammonia assimilation pathways in Rhodopseudomonas capsulata ElFl by Moreno-Vivian C. (1983)
  23. Orr , J. , and R. Haselkorn . 1981 . Kinetic and inhibition studies of glutamine synthetase from the cyanobacterium Anabaena 7120 . J. Biol. Chem. 256 : 13099 - 13104 . (10.1016/S0021-9258(18)43012-8) / J. Biol. Chem. / Kinetic and inhibition studies of glutamine synthetase from the cyanobacterium Anabaena 7120 by Orr J. (1981)
  24. Sakhno , 0. N. , R. N. Ivanovskii , and E. N. Kondrat'eva . 1981 . The glutamine synthetase-glutamate synthase system in Rhodopseudomonas sphaeroides . Mikrobiologiya 50 : 607 - 612 . / Mikrobiologiya / The glutamine synthetase-glutamate synthase system in Rhodopseudomonas sphaeroides by Sakhno 0. (1981)
  25. Schutt , H. , and H. Holzer . 1972 . Biological function of the ammonia-induced inactivation of glutamine synthetase in Escherichia coli . Eur. J. Biochem. 26 : 68 - 72 . (10.1111/j.1432-1033.1972.tb01740.x) / Eur. J. Biochem. / Biological function of the ammonia-induced inactivation of glutamine synthetase in Escherichia coli by Schutt H. (1972)
  26. Scolnick , P. A. , J. Virosco , and R. Haselkorn . 1983 . The wild-type gene for glutamine synthetase restores ammonia control of nitrogen fixation to Gln- (glnA) mutants of Rhodopseudomonas capsulata . J. Bacteriol. 155 : 180 - 185 . (10.1128/JB.155.1.180-185.1983) / J. Bacteriol. / The wild-type gene for glutamine synthetase restores ammonia control of nitrogen fixation to Gln- (glnA) mutants of Rhodopseudomonas capsulata by Scolnick P. A. (1983)
  27. Shapiro , B. M. , and E. R. Stadtman . 1970 . Glutamine synthetase (Escherichia coli) . Methods Enzymol. 17A : 910 - 922 . (10.1016/0076-6879(71)17305-3) / Methods Enzymol. / Glutamine synthetase (Escherichia coli) by Shapiro B. M. (1970)
  28. Siegel , L. M. , and K. J. Monty . 1966 . Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductase . Biochim. Biophys. Acta 112 : 346 - 362 . (10.1016/0926-6585(66)90333-5) / Biochim. Biophys. Acta / Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductase by Siegel L. M. (1966)
  29. Soliman , A. , S. Nordlund , B. C. Johansson , and H. Baltscheffsky . 1981 . Purification of glutamine synthetase from Rhodospirillum rubrum by affinity chromatography . Acta Chem.- Scand. B35 : 63 - 64 . (10.3891/acta.chem.scand.35b-0063) / Acta Chem.- Scand. / Purification of glutamine synthetase from Rhodospirillum rubrum by affinity chromatography by Soliman A. (1981)
  30. Stacey , G. , C. van Baalen , and F. R. Tabita . 1979 . Nitrogen and ammonia assimilation in the cyanobacteria: regulation of glutamine synthetase . Arch. Biochem. Biophys. 194 : 457 - 467 . (10.1016/0003-9861(79)90640-4) / Arch. Biochem. Biophys. / Nitrogen and ammonia assimilation in the cyanobacteria: regulation of glutamine synthetase by Stacey G. (1979)
  31. Stadtman E. R. and A. Ginsburg. 1974. The glutamine synthetase of Escherichia coli: structure and control p. 755-807. In P. D. Boyer (ed.) The enzymes vol. X 3rd ed. Academic Press Inc. New York. (10.1016/S1874-6047(08)60156-0)
  32. Stadtman , E. R. , A. Ginsburg , J. E. Ciardi , J. Yeh , S. B. Hennig , and B. M. Shapiro . 1970 . Multiple molecular forms of glutamine synthetase produced by enzyme-catalysed adenylylation and deadenylylation reactions . Adv. Enzyme Reg. 8 : 99 - 118 . (10.1016/0065-2571(70)90011-7) / Adv. Enzyme Reg. / Multiple molecular forms of glutamine synthetase produced by enzyme-catalysed adenylylation and deadenylylation reactions by Stadtman E. R. (1970)
  33. Stadtman , E. R. , P. Z. Smyrniotis , J. N. Davis , and M. E. Wittenberger . 1979 . Enzymic procedures for determining the average state of adenylylation of Escherichia coli glutamine synthetase . Anal. Biochem. 95 : 275 - 285 . (10.1016/0003-2697(79)90217-3) / Anal. Biochem. / Enzymic procedures for determining the average state of adenylylation of Escherichia coli glutamine synthetase by Stadtman E. R. (1979)
  34. Stewart G. R. and D. Rhodes. 1977. Control of enzyme levels in the regulation of nitrogen assimilation p. 1-19. In H. Smith (ed.) Regulation of enzyme synthesis and activity in higher plants. Academic Press Inc. New York.
  35. Weaver , P. F. , J. D. Wall , and H. Gest . 1975 . Characterization of Rhodopseudomonas capsulata . Arch. Microbiol. 105 : 207 - 216 . (10.1007/BF00447139) / Arch. Microbiol. / Characterization of Rhodopseudomonas capsulata by Weaver P. F. (1975)
  36. Weber , K. , and M. J. Osborn . 1969 . The reliability for molecular weight determination by dodecyl sulfate-polyacrylamide gel electrophoresis . J. Biol. Chem. 244 : 4406 - 4412 . (10.1016/S0021-9258(18)94333-4) / J. Biol. Chem. / The reliability for molecular weight determination by dodecyl sulfate-polyacrylamide gel electrophoresis by Weber K. (1969)
  37. Wedler , F. C. , R. M. Kenney , A. E. Ashour , and J. Carfi . 1978 . Two regulatory isozymes of glutamine synthetase from Bacillus caldolyticus, an extreme thermophile . Biochem. Biophys. Res. Commun. 81 : 122 - 126 . (10.1016/0006-291X(78)91638-8) / Biochem. Biophys. Res. Commun. / Two regulatory isozymes of glutamine synthetase from Bacillus caldolyticus, an extreme thermophile by Wedler F. C. (1978)
  38. Wohlhueter R. M. H. Schutt and H. Holzer. 1973. Regulation of glutamine synthesis in vivo in E. coli p. 45-64. In S. Prusiner and E. R. Stadtman (ed.) The enzymes of glutamine metabolism. Academic Press Inc. New York.
  39. Woolfolk , C. A. , and E. R. Stadtman . 1967 . Regulation of glutamine synthetase. III. Cumulative feedback inhibition of glutamine synthetase from Escherichia coli . Arch. Biochem. Biophys. 118 : 736 - 755 . / Arch. Biochem. Biophys. / Regulation of glutamine synthetase. III. Cumulative feedback inhibition of glutamine synthetase from Escherichia coli by Woolfolk C. A. (1967)
  40. Yoch , D. C. , M. Cantu , and Z. -M. Zhang . 1983 . Evidence for a glutamine synthetase-chromatophore association in the phototroph Rhodospirillum rubrum: purification, properties, and regulation of the enzyme . J. Bacteriol. 154 : 632 - 639 . (10.1128/jb.154.2.632-639.1983) / J. Bacteriol. / Evidence for a glutamine synthetase-chromatophore association in the phototroph Rhodospirillum rubrum: purification, properties, and regulation of the enzyme by Yoch D. C. (1983)
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Created 5 years, 8 months ago (Jan. 3, 2020, 1:08 p.m.)
Deposited 4 years, 3 months ago (June 1, 2021, 1:02 p.m.)
Indexed 1 year, 9 months ago (Nov. 23, 2023, 11:23 a.m.)
Issued 40 years, 4 months ago (May 1, 1985)
Published 40 years, 4 months ago (May 1, 1985)
Published Print 40 years, 4 months ago (May 1, 1985)
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@article{Caballero_1985, title={Molecular and regulatory properties of glutamine synthetase from the phototrophic bacterium Rhodopseudomonas capsulata E1F1}, volume={162}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.162.2.804-809.1985}, DOI={10.1128/jb.162.2.804-809.1985}, number={2}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Caballero, F J and Cejudo, F J and Florencio, F J and Cárdenas, J and Castillo, F}, year={1985}, month=may, pages={804–809} }