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. 1976 Jun;126(3):1271–1277. doi: 10.1128/jb.126.3.1271-1277.1976

Molecular and catalytic properties of ribulose 1,5-bisphosphate carboxylase from the photosynthetic extreme halophile Ectothiorhodospira halophila.

F R Tabita, B A McFadden
PMCID: PMC233153  PMID: 947890

Abstract

D-Ribulose 1,5-bisphosphate (RuBP) carboxylase has been purified from the photosynthetic extreme halophile Ectothiorhodospira halophila. Despite a growth requirement for almost saturating sodium chloride in the medium, both crude and homogeneous preparations of RuBP carboxylase obtained from this organism were inhibited by salts. Sedimentation equilibrium analyses showed the enzyme to be large (molecular weight: 601,000). The protein was composed of two types of polypeptide chains of 56,000 and of 18,000 daltons. The small subunit appeared to be considerably larger than the small subunit obtained from the RuBP carboxylase isolated from Chromatium, an organism related to E. halophila. Amino acid analyses of hydrolysates of both E. halophilia and Chromatium RuBP carboxylases were very similar. Initial velocity experiments showed that the E. halophila RuBP carboxylase had a Km for ribulose diphosphate of 0.07 mM and a Km for HCO3- of 10 mM. Moreover, 6-phospho-D-gluconate was found to markedly inhibit the E. halophila carboxylase; a Ki for phosphogluconate of 0.14 mM was determined.

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Selected References

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

  1. Akazawa T., Kondo H., Shimazue T., Nishimura M., Sugiyama T. Further studies on ribulose 1,5-diphosphate carboxylase from Chromatium strain D. Biochemistry. 1972 Mar 28;11(7):1298–1303. doi: 10.1021/bi00757a028. [DOI] [PubMed] [Google Scholar]
  2. CHRISTIAN J. H., WALTHO J. A. Solute concentrations within cells of halophilic and non-halophilic bacteria. Biochim Biophys Acta. 1962 Dec 17;65:506–508. doi: 10.1016/0006-3002(62)90453-5. [DOI] [PubMed] [Google Scholar]
  3. Chu D. K., Bassham J. A. Activation and inhibition of ribulose 1,5-diphosphate carboxylase by 6-phosphogluconate. Plant Physiol. 1973 Oct;52(4):373–379. doi: 10.1104/pp.52.4.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HIRS C. H. The oxidation of ribonuclease with performic acid. J Biol Chem. 1956 Apr;219(2):611–621. [PubMed] [Google Scholar]
  5. Kuehn G. D., McFadden B. A. Ribulose 1,5-diphosphate carboxylase from Hydrogenomonas eutropha and Hydrogenomonas facilis. I. Purification, metallic ion requirements, inhibition, and kinetic constants. Biochemistry. 1969 Jun;8(6):2394–2402. doi: 10.1021/bi00834a021. [DOI] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. McFadden B. A. Autotrophic CO2 assimilation and the evolution of ribulose diphosphate carboxylase. Bacteriol Rev. 1973 Sep;37(3):289–319. doi: 10.1128/br.37.3.289-319.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. McFadden B. A., Tabita F. R. D-ribulose-1, 5-diphosphate carboxylase and the evolution of autotrophy. Biosystems. 1974 Oct;6(2):93–112. doi: 10.1016/0303-2647(74)90002-1. [DOI] [PubMed] [Google Scholar]
  9. McFadden B. A., Tu C. C. Regulation of autotrophic and heterotrophic carbon dioxide fixation in Hydrogenomonas facilis. J Bacteriol. 1967 Mar;93(3):886–893. doi: 10.1128/jb.93.3.886-893.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nishimura M., Akazawa T. Further proof for the catalytic role of the larger subunit in the spinach leaf ribulose-1,5-diphosphate carboxylase. Biochem Biophys Res Commun. 1973 Oct 1;54(3):842–848. doi: 10.1016/0006-291x(73)90770-5. [DOI] [PubMed] [Google Scholar]
  11. Nishimura M., Akazawa T. Reconstitution of spinach ribulose-1,5-diphosphate carboxylase from departed subunits. Biochem Biophys Res Commun. 1974 Jul 24;59(2):584–590. doi: 10.1016/s0006-291x(74)80020-3. [DOI] [PubMed] [Google Scholar]
  12. Raymond J. C., Sistrom W. R. The isolation and preliminary characterization of a halophilic photosynthetic bacterium. Arch Mikrobiol. 1967;59(1):255–268. doi: 10.1007/BF00406339. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Tabita F. R., McFadden B. A. One-step isolation of microbial ribulose-1,5-diphosphate carboxylase. Arch Microbiol. 1974;99(3):231–240. doi: 10.1007/BF00696237. [DOI] [PubMed] [Google Scholar]
  15. Tabita F. R., McFadden B. A., Pfennig N. D-ribulose-1,5-bisphosphate carboxylase in Chlorobium thiosulfatophilum Tassajara. Biochim Biophys Acta. 1974 Mar 21;341(1):187–194. doi: 10.1016/0005-2744(74)90079-5. [DOI] [PubMed] [Google Scholar]
  16. Tabita F. R., McFadden B. A. Regulation of ribulose-1,5-diphosphate carboxylase by 6-phospho-D-gluconate. Biochem Biophys Res Commun. 1972 Sep 5;48(5):1153–1159. doi: 10.1016/0006-291x(72)90831-5. [DOI] [PubMed] [Google Scholar]
  17. Tabita F. R., Stevens S. E., Jr, Gibson J. L. Carbon dioxide assimilation in blue-green algae: initial studies on the structure of ribulose 1,5-bisphosphate carboxylase. J Bacteriol. 1976 Feb;125(2):531–539. doi: 10.1128/jb.125.2.531-539.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tabita R. F., Stevens S. E., Jr, Quijano R. D-ribulose 1, 5-diphosphate carboxylase from blue-green algae. Biochem Biophys Res Commun. 1974 Nov 6;61(1):45–52. doi: 10.1016/0006-291x(74)90531-2. [DOI] [PubMed] [Google Scholar]
  19. Takabe T., Akazawa T. Catalytic role of subunit A in ribulose-1,5-diphosphate carboxylase from Chromatium strain D. Arch Biochem Biophys. 1973 Jul;157(1):303–308. doi: 10.1016/0003-9861(73)90415-3. [DOI] [PubMed] [Google Scholar]
  20. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]
  21. Wishnick M., Lane M. D., Scrutton M. C. The interaction of metal ions with ribulose 1,5-diphosphate carboxylase from spinach. J Biol Chem. 1970 Oct 10;245(19):4939–4947. [PubMed] [Google Scholar]
  22. YPHANTIS D. A. EQUILIBRIUM ULTRACENTRIFUGATION OF DILUTE SOLUTIONS. Biochemistry. 1964 Mar;3:297–317. doi: 10.1021/bi00891a003. [DOI] [PubMed] [Google Scholar]

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