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. 1981 Jun;67(6):1220–1223. doi: 10.1104/pp.67.6.1220

Storage and Maintaining Activity of Ribulose Bisphosphate Carboxylase/Oxygenase 1

Nigel P Hall 1,2, Stephen D McCurry 1,3, N E Tolbert 1
PMCID: PMC425865  PMID: 16661840

Abstract

Purified ribulose-1,5-bisphosphate carboxylase/oxygenase in 50% saturated (NH4)2SO4 was stable when frozen as small beads in liquid nitrogen and stored at −80 C. When stored as a slurry at 4 C most of the activity was lost within four weeks. This loss was due not only to enzyme polymerization. Activity in old preparations purified from spinach leaves, but not tobacco or tomato leaves, can be restored to the level of newly purified enzyme after storage at 4 C by treatment with 50 to 100 millimolar dithiothreitol for several hours followed by dialysis against buffer and 1 millimolar dithiothreitol before CO2 and Mg2+ activation and assay. Some enzyme oligomers that had been formed were not converted back to native enzyme by treatment with 100 millimolar dithiothreitol.

The purified enzyme contained about 2 gram-atoms iron per mole enzyme that could not be removed by chelating agents. When the enzyme was incubated with 100 millimolar dithiothreitol and exposed to O2, a purple dithiothreitol-iron complex was formed which could be removed by dialysis. The activities of ribulose-1,5-bisphosphate carboxylase and oxygenase were not altered by reducing the iron content to 0.7 mole per mole enzyme by treatment with dithiothreitol followed by exhaustive dialysis against iron free buffer.

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Selected References

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

  1. Bentle L. A., Lardy H. A. P-enolpyruvate carboxykinase ferroactivator. Purification and some properties. J Biol Chem. 1977 Feb 25;252(4):1431–1440. [PubMed] [Google Scholar]
  2. CLELAND W. W. DITHIOTHREITOL, A NEW PROTECTIVE REAGENT FOR SH GROUPS. Biochemistry. 1964 Apr;3:480–482. doi: 10.1021/bi00892a002. [DOI] [PubMed] [Google Scholar]
  3. Chan P. H., Sakano K., Singh S., Wildman S. G. Crystalline fraction I protein: preparation in large yield. Science. 1972 Jun 9;176(4039):1145–1146. doi: 10.1126/science.176.4039.1145. [DOI] [PubMed] [Google Scholar]
  4. Chollet R., Anderson L. L., Hovsepian L. C. The absence of tightly bound copper, iron, and flavin nucleotide in crystalline ribulose 1,5-bisphosphate carboxylase-oxygenase from tobacco. Biochem Biophys Res Commun. 1975 May 5;64(1):97–107. doi: 10.1016/0006-291x(75)90224-7. [DOI] [PubMed] [Google Scholar]
  5. Cybulsky D. L., Nagy A., Kandel S. I., Kandel M., Gornall A. G. Carbonic anhydrase from spinach leaves. Chemical modification and affinity labeling. J Biol Chem. 1979 Mar 25;254(6):2032–2039. [PubMed] [Google Scholar]
  6. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  7. Johal S., Bourque D. P. Crystalline ribulose 1,5-bisphosphate carboxylase-oxygenase from spinach. Science. 1979 Apr 6;204(4388):75–77. doi: 10.1126/science.204.4388.75. [DOI] [PubMed] [Google Scholar]
  8. Lorimer G. H., Andrews T. J., Tolbert N. E. Ribulose diphosphate oxygenase. II. Further proof of reaction products and mechanism of action. Biochemistry. 1973 Jan 2;12(1):18–23. doi: 10.1021/bi00725a004. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. McCurry S. D., Hall N. P., Pierce J., Paech C., Tolbert N. E. Ribulose-1,5-bisphosphate carboxylase/oxygenase from parsley. Biochem Biophys Res Commun. 1978 Oct 30;84(4):896–900. doi: 10.1016/0006-291x(78)91667-4. [DOI] [PubMed] [Google Scholar]
  11. McCurry S. D., Tolbert N. E. Inhibition of ribulose-1,5-bisphosphate carboxylase/oxygenase by xylulose 1,5-bisphosphate. J Biol Chem. 1977 Dec 10;252(23):8344–8346. [PubMed] [Google Scholar]
  12. 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]
  13. Ryan F. J., Barker R., Tolbert N. E. Inhibition of ribulose diphosphate carboxylase/oxygenase by xylitol 1,5-diphosphate. Biochem Biophys Res Commun. 1975 Jul 8;65(1):39–46. doi: 10.1016/s0006-291x(75)80058-1. [DOI] [PubMed] [Google Scholar]
  14. Ryan F. J., Tolbert N. E. Ribulose diphosphate carboxylase/oxygenase. III. Isolation and properties. J Biol Chem. 1975 Jun 10;250(11):4229–4233. [PubMed] [Google Scholar]
  15. Schloss J. V., Phares E. F., Long M. V., Norton I. L., Stringer C. D., Hartman F. C. Isolation, characterization, and crystallization of ribulosebisphosphate carboxylase from autotrophically grown Rhodospirillum rubrum. J Bacteriol. 1979 Jan;137(1):490–501. doi: 10.1128/jb.137.1.490-501.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Van de Bogart M., Beinert H. Micro methods for the quantitative determination of iron and copper in biological material. Anal Biochem. 1967 Aug;20(2):325–334. doi: 10.1016/0003-2697(67)90038-3. [DOI] [PubMed] [Google Scholar]
  17. Wishnick M., Lane M. D., Scrutton M. C., Mildvan A. S. The presence of tightly bound copper in ribulose diphosphate carboxylase from spinach. J Biol Chem. 1969 Oct 25;244(20):5761–5763. [PubMed] [Google Scholar]

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