Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Nov 1;287(Pt 3):821–825. doi: 10.1042/bj2870821

Purification and properties of a phosphatase in French bean (Phaseolus vulgaris L.) leaves that hydrolyses 2'-carboxy-D-arabinitol 1-phosphate.

A H Kingston-Smith 1, I Major 1, M A Parry 1, A J Keys 1
PMCID: PMC1133081  PMID: 1332682

Abstract

An enzyme that releases P(i) from 2-carboxy-D-arabinitol 1-phosphate, a naturally occurring tightly binding inhibitor of ribulose 1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), was purified from leaves of French bean seedlings. It was a monomeric protein of M(r) about 56,000. Catalytic activity was stimulated by increased concentrations of inorganic salts to a maximum at an ionic strength above 0.2. NADPH and D-fructose 1,6-bisphosphate increased the activity of the enzyme in both the presence and absence of 0.2 M-KCl. The pure enzyme did not require dithiothreitol for activity. The pH optimum was 7, the Km for 2-carboxy-D-arabinitol 1-phosphate was 0.43 mM and the specific activity 6.8 mumol/min per mg of protein. The enzyme had little or no activity against phosphate ester intermediates of photosynthetic metabolism and glycolysis but hydrolysed the 1,5-bisphosphates of 2'-carboxy-D-ribitol and 2'-carboxy-D-arabinitol more rapidly than 2'-carboxy-D-arabinitol 1-phosphate.

Full text

PDF
821

Images in this article

822Fig. 1.
on p.822

Selected References

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

  1. Beck E., Scheibe R., Reiner J. An Assessment of the Rubisco Inhibitor: 2-Carboxyarabinitol-1-Phosphate and d-Hamamelonic Acid 2-Phosphate Are Identical Compounds. Plant Physiol. 1989 May;90(1):13–16. doi: 10.1104/pp.90.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beck E., Stransky H., Fürbringer M. Synthesis of hamamelose-diphosphate by isolated spinach chloroplasts. FEBS Lett. 1971 Mar 16;13(4):229–234. doi: 10.1016/0014-5793(71)80542-2. [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. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Gutteridge S., Reddy G. S., Lorimer G. The synthesis and purification of 2'-carboxy-D-arabinitol 1-phosphate, a natural inhibitor of ribulose 1,5-bisphosphate carboxylase, investigated by 31P n.m.r. Biochem J. 1989 Jun 15;260(3):711–716. doi: 10.1042/bj2600711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Holbrook G. P., Bowes G., Salvucci M. E. Degradation of 2-carboxyarabinitol 1-phosphate by a specific chloroplast phosphatase. Plant Physiol. 1989 Jun;90(2):673–678. doi: 10.1104/pp.90.2.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Holbrook G. P., Galasinski S. C., Salvucci M. E. Regulation of 2-carboxyarabinitol 1-phosphatase. Plant Physiol. 1991 Nov;97(3):894–899. doi: 10.1104/pp.97.3.894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Husic H. D., Tolbert N. E. Anion and divalent cation activation of phosphoglycolate phosphatase from leaves. Arch Biochem Biophys. 1984 Feb 15;229(1):64–72. doi: 10.1016/0003-9861(84)90130-9. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Pierce J., Tolbert N. E., Barker R. Interaction of ribulosebisphosphate carboxylase/oxygenase with transition-state analogues. Biochemistry. 1980 Mar 4;19(5):934–942. doi: 10.1021/bi00546a018. [DOI] [PubMed] [Google Scholar]
  11. Salvucci M. E., Holbrook G. P. Purification and Properties of 2-Carboxy-d-Arabinitol 1-Phosphatase. Plant Physiol. 1989 Jun;90(2):679–685. doi: 10.1104/pp.90.2.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Van Veldhoven P. P., Mannaerts G. P. Inorganic and organic phosphate measurements in the nanomolar range. Anal Biochem. 1987 Feb 15;161(1):45–48. doi: 10.1016/0003-2697(87)90649-x. [DOI] [PubMed] [Google Scholar]
  13. Vu C. V., Allen L. H., Bowes G. Effects of Light and Elevated Atmospheric CO(2) on the Ribulose Bisphosphate Carboxylase Activity and Ribulose Bisphosphate Level of Soybean Leaves. Plant Physiol. 1983 Nov;73(3):729–734. doi: 10.1104/pp.73.3.729. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES