Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Jun;174(11):3593–3600. doi: 10.1128/jb.174.11.3593-3600.1992

Reversible inactivation and characterization of purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides.

X Wang 1, F R Tabita 1
PMCID: PMC206046  PMID: 1592814

Abstract

Form I ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) from Rhodobacter sphaeroides is inactivated upon the addition of organic acids to photolithoautotrophically grown cultures. Activity recovers after the dissipation of the organic acid from the culture. The inactivation process depends on both the concentration of the organic compound and the nitrogen status of the cells. The inactivated RubisCO has been purified and was shown to exhibit mobility on both nondenaturing and sodium dodecyl sulfate gels different from that of the active enzyme prepared from cells not treated with organic acids. However, the Michaelis constants for ribulose 1,5-bisphosphate and CO2 or O2 were not dramatically altered. Purified inactivated RubisCO could be activated in vitro by increasing the temperature or the levels of Mg(II), and this activation was accompanied by changes in the electrophoretic mobility of the protein. When foreign bacterial RubisCO genes were expressed in an R. sphaeroides host strain lacking the ability to synthesize endogenous RubisCO, only slight inactivation of RubisCO activity was attained.

Full text

PDF
3599

Images in this article

3597Image
on p.3597

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersen K., Wilke-Douglas M. Genetic and physical mapping and expression in Pseudomonas aeruginosa of the chromosomally encoded ribulose bisphosphate carboxylase genes of Alcaligenes eutrophus. J Bacteriol. 1987 May;169(5):1997–2004. doi: 10.1128/jb.169.5.1997-2004.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arp D. J., Zumft W. G. L-methionine-SR-sulfoximine as a probe for the role of glutamine synthetase in nitrogenase switch-off by ammonia and glutamine in Rhodopseudomonas palustris. Arch Microbiol. 1983 Jan;134(1):17–22. doi: 10.1007/BF00429400. [DOI] [PubMed] [Google Scholar]
  3. Berry J. A., Lorimer G. H., Pierce J., Seemann J. R., Meek J., Freas S. Isolation, identification, and synthesis of 2-carboxyarabinitol 1-phosphate, a diurnal regulator of ribulose-bisphosphate carboxylase activity. Proc Natl Acad Sci U S A. 1987 Feb;84(3):734–738. doi: 10.1073/pnas.84.3.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CHANEY A. L., MARBACH E. P. Modified reagents for determination of urea and ammonia. Clin Chem. 1962 Apr;8:130–132. [PubMed] [Google Scholar]
  5. Falcone D. L., Quivey R. G., Jr, Tabita F. R. Transposon mutagenesis and physiological analysis of strains containing inactivated form I and form II ribulose bisphosphate carboxylase/oxygenase genes in Rhodobacter sphaeroides. J Bacteriol. 1988 Jan;170(1):5–11. doi: 10.1128/jb.170.1.5-11.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Falcone D. L., Tabita F. R. Expression of endogenous and foreign ribulose 1,5-bisphosphate carboxylase-oxygenase (RubisCO) genes in a RubisCO deletion mutant of Rhodobacter sphaeroides. J Bacteriol. 1991 Mar;173(6):2099–2108. doi: 10.1128/jb.173.6.2099-2108.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fitzmaurice W. P., Saari L. L., Lowery R. G., Ludden P. W., Roberts G. P. Genes coding for the reversible ADP-ribosylation system of dinitrogenase reductase from Rhodospirillum rubrum. Mol Gen Genet. 1989 Aug;218(2):340–347. doi: 10.1007/BF00331287. [DOI] [PubMed] [Google Scholar]
  8. Gibson J. L., Chen J. H., Tower P. A., Tabita F. R. The form II fructose 1,6-bisphosphatase and phosphoribulokinase genes form part of a large operon in Rhodobacter sphaeroides: primary structure and insertional mutagenesis analysis. Biochemistry. 1990 Sep 4;29(35):8085–8093. doi: 10.1021/bi00487a014. [DOI] [PubMed] [Google Scholar]
  9. Gibson J. L., Falcone D. L., Tabita F. R. Nucleotide sequence, transcriptional analysis, and expression of genes encoded within the form I CO2 fixation operon of Rhodobacter sphaeroides. J Biol Chem. 1991 Aug 5;266(22):14646–14653. [PubMed] [Google Scholar]
  10. Gibson J. L., Tabita F. R. Characterization of antiserum directed against form II ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides. J Bacteriol. 1977 Sep;131(3):1020–1022. doi: 10.1128/jb.131.3.1020-1022.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gibson J. L., Tabita F. R. Different molecular forms of D-ribulose-1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides. J Biol Chem. 1977 Feb 10;252(3):943–949. [PubMed] [Google Scholar]
  12. Gibson J. L., Tabita F. R. Structural differences in the catalytic subunits of form I and form II ribulose 1,5-bisphosphate carboxylase/oxygenase from Rhodopseudomonas sphaeroides. J Bacteriol. 1985 Dec;164(3):1188–1193. doi: 10.1128/jb.164.3.1188-1193.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HURLBERT R. E., LASCELLES J. RIBULOSE DIPHOSPHATE CARBOXYLASE IN THIORHODACEAE. J Gen Microbiol. 1963 Dec;33:445–458. doi: 10.1099/00221287-33-3-445. [DOI] [PubMed] [Google Scholar]
  14. Hallenbeck P. L., Lerchen R., Hessler P., Kaplan S. Phosphoribulokinase activity and regulation of CO2 fixation critical for photosynthetic growth of Rhodobacter sphaeroides. J Bacteriol. 1990 Apr;172(4):1749–1761. doi: 10.1128/jb.172.4.1749-1761.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jouanneau Y., Tabita F. R. In vivo regulation of form I ribulose 1,5-bisphosphate carboxylase/oxygenase from Rhodopseudomonas sphaeroides. Arch Biochem Biophys. 1987 Apr;254(1):290–303. doi: 10.1016/0003-9861(87)90105-6. [DOI] [PubMed] [Google Scholar]
  16. Jouanneau Y., Tabita F. R. Independent regulation of synthesis of form I and form II ribulose bisphosphate carboxylase-oxygenase in Rhodopseudomonas sphaeroides. J Bacteriol. 1986 Feb;165(2):620–624. doi: 10.1128/jb.165.2.620-624.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuehn G. D., McFadden B. A. Factors affecting the synthesis and degradation of ribulose-1,5-diphosphate carboxylase in Hydrogenomonas facilis and Hydrogenomonas eutropha. J Bacteriol. 1968 Mar;95(3):937–946. doi: 10.1128/jb.95.3.937-946.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee B., Tabita F. R. Purification of recombinant ribulose-1,5-bisphosphate carboxylase/oxygenase large subunits suitable for reconstitution and assembly of active L8S8 enzyme. Biochemistry. 1990 Oct 9;29(40):9352–9357. doi: 10.1021/bi00492a007. [DOI] [PubMed] [Google Scholar]
  19. Lowery R. G., Ludden P. W. Purification and properties of dinitrogenase reductase ADP-ribosyltransferase from the photosynthetic bacterium Rhodospirillum rubrum. J Biol Chem. 1988 Nov 15;263(32):16714–16719. [PubMed] [Google Scholar]
  20. Lowery R. G., Saari L. L., Ludden P. W. Reversible regulation of the nitrogenase iron protein from Rhodospirillum rubrum by ADP-ribosylation in vitro. J Bacteriol. 1986 May;166(2):513–518. doi: 10.1128/jb.166.2.513-518.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mann N. H., Turner A. M. Covalent modification of ribulose 1,5-bisphosphate carboxylase/oxygenase in Rhodomicrobium vannielii. Mol Microbiol. 1988 May;2(3):427–432. doi: 10.1111/j.1365-2958.1988.tb00048.x. [DOI] [PubMed] [Google Scholar]
  22. Meijer W. G., Arnberg A. C., Enequist H. G., Terpstra P., Lidstrom M. E., Dijkhuizen L. Identification and organization of carbon dioxide fixation genes in Xanthobacter flavus H4-14. Mol Gen Genet. 1991 Feb;225(2):320–330. doi: 10.1007/BF00269865. [DOI] [PubMed] [Google Scholar]
  23. ORMEROD J. G., ORMEROD K. S., GEST H. Light-dependent utilization of organic compounds and photoproduction of molecular hydrogen by photosynthetic bacteria; relationships with nitrogen metabolism. Arch Biochem Biophys. 1961 Sep;94:449–463. doi: 10.1016/0003-9861(61)90073-x. [DOI] [PubMed] [Google Scholar]
  24. Read B. A., Tabita F. R. Amino acid substitutions in the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase that influence catalytic activity of the holoenzyme. Biochemistry. 1992 Jan 21;31(2):519–525. doi: 10.1021/bi00117a031. [DOI] [PubMed] [Google Scholar]
  25. Saari L. L., Pope M. R., Murrell S. A., Ludden P. W. Studies on the activating enzyme for iron protein of nitrogenase from Rhodospirillum rubrum. J Biol Chem. 1986 Apr 15;261(11):4973–4977. [PubMed] [Google Scholar]
  26. Sarles L. S., Tabita F. R. Derepression of the synthesis of D-ribulose 1,5-bisphosphate carboxylase/oxygenase from Rhodospirillum rubrum. J Bacteriol. 1983 Jan;153(1):458–464. doi: 10.1128/jb.153.1.458-464.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shively J. M., Davidson E., Marrs B. L. Depression of the synthesis of the intermediate and large forms of ribulose-1,5-bisphosphate carboxylase/oxygenase in Rhodopseudomonas capsulata. Arch Microbiol. 1984 Jul;138(3):233–236. doi: 10.1007/BF00402127. [DOI] [PubMed] [Google Scholar]
  28. Tabita F. R., McFadden B. A. D-ribulose 1,5-diphosphate carboxylase from Rhodospirillum rubrum. I. Levels, purification, and effects of metallic ions. J Biol Chem. 1974 Jun 10;249(11):3453–3458. [PubMed] [Google Scholar]
  29. Tabita F. R. Molecular and cellular regulation of autotrophic carbon dioxide fixation in microorganisms. Microbiol Rev. 1988 Jun;52(2):155–189. doi: 10.1128/mr.52.2.155-189.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang X., Tabita F. R. Interaction between ribulose 1,5-bisphosphate carboxylase/oxygenase activity and the ammonia assimilatory system of Rhodobacter sphaeroides. J Bacteriol. 1992 Jun;174(11):3601–3606. doi: 10.1128/jb.174.11.3601-3606.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weaver K. E., Tabita F. R. Isolation and partial characterization of Rhodopseudomonas sphaeroides mutants defective in the regulation of ribulose bisphosphate carboxylase/oxygenase. J Bacteriol. 1983 Nov;156(2):507–515. doi: 10.1128/jb.156.2.507-515.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Werneke J. M., Chatfield J. M., Ogren W. L. Catalysis of Ribulosebisphosphate Carboxylase/Oxygenase Activation by the Product of a Rubisco Activase cDNA Clone Expressed in Escherichia coli. Plant Physiol. 1988 Aug;87(4):917–920. doi: 10.1104/pp.87.4.917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Whitman W., Tabita F. R. Inhibition of D-ribulose 1,5-bisphosphate carboxylase by pyridoxal 5'-phosphate. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1034–1039. doi: 10.1016/0006-291x(76)90758-0. [DOI] [PubMed] [Google Scholar]
  34. Yoch D. C., Cantu M. Changes in the regulatory form of Rhodospirillum rubrum nitrogenase as influenced by nutritional and environmental factors. J Bacteriol. 1980 Jun;142(3):899–907. doi: 10.1128/jb.142.3.899-907.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES