Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1987 Jun;84(2):483–490. doi: 10.1104/pp.84.2.483

Structural, Functional, and Evolutionary Analysis of Ribulose-1,5-Bisphosphate Carboxylase from the Chromophytic Alga Olisthodiscus luteus1

Scott M Newman 1, Rose Ann Cattolico 1
PMCID: PMC1056607  PMID: 16665466

Abstract

Ribulose-1,5-bisphosphate carboxylase (RuBPCase) was purified from the marine chromophyte Olisthodiscus luteus. This study represents the first extensive analysis of RuBPCase from a chromophytic plant species as well as from an organism where both subunits of the enzyme are encoded on the chloroplast genome. The size of the purified holoenzyme (17.9 Svedberg units, 588 kilodaltons) was determined by sedimentation analysis and the size of the subunits (55 kilodaltons, 15 kilodaltons) ascertained by analytical sodium dodecyl sulfate gel electrophoresis. This data predicts either an 8:9 or 8:8 ratio of the large to small subunits in the holoenzyme. Amino acid analyses demonstrate that the O. luteus RuBPCase large subunit is highly conserved and the small subunit much less so when compared with the chlorophytic plant peptides. The catalytic optima of pH and Mg2+ have been determined as well as the response of enzyme catalysis to temperature. The requirements of NaHCO3 and Mg2+ for enzyme activation have also been analyzed. The Michaelis constants for the substrates of the carboxylation reaction (CO2 and ribulose bisphosphate) were shown to be 45 and 48 micromolar, respectively. Competitive inhibition by oxygen of RuBPCase-catalyzed CO2 fixation was also demonstrated. These data demonstrate that a high degree of RuBPCase conservation occurs among widely divergent photoautotrophs regardless of small subunit coding site.

Full text

PDF
483

Images in this article

486Fig. 2
on p.486

Selected References

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

  1. Andrews T. J., Abel K. M. Kinetics and subunit interactions of ribulose bisphosphate carboxylase-oxygenase from the cyanobacterium, Synechococcus sp. J Biol Chem. 1981 Aug 25;256(16):8445–8451. [PubMed] [Google Scholar]
  2. Bowien B., Mayer F. Further studies on the quaternary structure of D-ribulose-1, 5-bisphosphate carboxylase from Alcaligenes eutrophus. Eur J Biochem. 1978 Jul 17;88(1):97–107. doi: 10.1111/j.1432-1033.1978.tb12426.x. [DOI] [PubMed] [Google Scholar]
  3. Ford T. W. Ribulose 1,5-bisphosphate carboxylase from the thermophilic, acidophilic alga, Cyanidium caldarium (Geitler). Purification, characterisation and thermostability of the enzyme. Biochim Biophys Acta. 1979 Aug 15;569(2):239–248. doi: 10.1016/0005-2744(79)90059-7. [DOI] [PubMed] [Google Scholar]
  4. Jordan D. B., Ogren W. L. Species variation in kinetic properties of ribulose 1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys. 1983 Dec;227(2):425–433. doi: 10.1016/0003-9861(83)90472-1. [DOI] [PubMed] [Google Scholar]
  5. Lorimer G. H., Badger M. R., Andrews T. J. The activation of ribulose-1,5-bisphosphate carboxylase by carbon dioxide and magnesium ions. Equilibria, kinetics, a suggested mechanism, and physiological implications. Biochemistry. 1976 Feb 10;15(3):529–536. doi: 10.1021/bi00648a012. [DOI] [PubMed] [Google Scholar]
  6. McCurry S. D., Gee R., Tolbert N. E. Ribulose-1,5-bisphosphate carboxylase/oxygenase from spinach, tomato, or tobacco leaves. Methods Enzymol. 1982;90(Pt E):515–521. doi: 10.1016/s0076-6879(82)90178-1. [DOI] [PubMed] [Google Scholar]
  7. McFadden B. A., Lord J. M., Rowe A., Dilks S. Composition, quaternary structure, and catalytic properties of D-ribulose-1, 5-bisphosphate carboxylase from Euglena gracilis. Eur J Biochem. 1975 May;54(1):195–206. doi: 10.1111/j.1432-1033.1975.tb04129.x. [DOI] [PubMed] [Google Scholar]
  8. Miziorko H. M., Lorimer G. H. Ribulose-1,5-bisphosphate carboxylase-oxygenase. Annu Rev Biochem. 1983;52:507–535. doi: 10.1146/annurev.bi.52.070183.002451. [DOI] [PubMed] [Google Scholar]
  9. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  10. Nierzwicki-Bauer S. A., Curtis S. E., Haselkorn R. Cotranscription of genes encoding the small and large subunits of ribulose-1,5-bisphosphate carboxylase in the cyanobacterium Anabaena 7120. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5961–5965. doi: 10.1073/pnas.81.19.5961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Paulsen J. M., Lane M. D. Spinach ribulose diphosphate carboxylase. I. Purification and properties of the enzyme. Biochemistry. 1966 Jul;5(7):2350–2357. doi: 10.1021/bi00871a025. [DOI] [PubMed] [Google Scholar]
  12. Plumley F. G., Kirchman D. L., Hodson R. E., Schmidt G. W. Ribulose Bisphosphate Carboxylase from Three Chlorophyll c-Containing Algae : Physical and Immunological Characterizations. Plant Physiol. 1986 Mar;80(3):685–691. doi: 10.1104/pp.80.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Reith M., Cattolico R. A. Inverted repeat of Olisthodiscus luteus chloroplast DNA contains genes for both subunits of ribulose-1,5-bisphosphate carboxylase and the 32,000-dalton Q(B) protein: Phylogenetic implications. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8599–8603. doi: 10.1073/pnas.83.22.8599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  15. Shinozaki K., Yamada C., Takahata N., Sugiura M. Molecular cloning and sequence analysis of the cyanobacterial gene for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4050–4054. doi: 10.1073/pnas.80.13.4050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sugiyama T., Ito T., Akazawa T. Subunit structure of ribulose 1,5-diphosphate carboxylase from Chlorella ellipsoidea. Biochemistry. 1971 Aug 31;10(18):3406–3411. doi: 10.1021/bi00794a014. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Whitman W. B., Martin M. N., Tabita F. R. Activation and regulation of ribulose bisphosphate carboxylase-oxygenase in the absence of small subunits. J Biol Chem. 1979 Oct 25;254(20):10184–10189. [PubMed] [Google Scholar]
  20. Yeoh H. H., Badger M. R., Watson L. Variations in Kinetic Properties of Ribulose-1,5-bisphosphate Carboxylases among Plants. Plant Physiol. 1981 Jun;67(6):1151–1155. doi: 10.1104/pp.67.6.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES