Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1970 Aug;46(2):204–207. doi: 10.1104/pp.46.2.204

A Simplified Purification and Some Properties of Ribulose 1,5-Diphosphate Carboxylase from Barley

G E Kleinkopf a, R C Huffaker a, A Matheson a
PMCID: PMC396563  PMID: 16657435

Abstract

A rapid procedure was developed for purifying ribulose 1,5-diphosphate carboxylase from barley leaves. After (NH4)2SO4 fractionation, the unique sedimentation properties of the enzyme were exploited to effect a single step purification to 90% homogeneity. High speed centrifugation pelleted the enzyme with complete recovery of activity. Residual impurities were then removed by diethylaminoethyl cellulose chromatography and density gradient centrifugation. The purified protein exhibited size heterogeneity due to polymerization. The polymerization products were enzymatically active aggregates of ribulose 1,5-diphosphate carboxylase and were precipitated by an antibody specific for the enzyme.

Full text

PDF

Images in this article

205Image
on p.205
205Image
on p.205
206Image
on p.206

Selected References

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

  1. Andersen W. R., Wildner G. F., Criddle R. S. Ribulose diphosphate carboxylase. 3. Altered forms of ribulose diphosphate carboxylase from mutant tomato plants. Arch Biochem Biophys. 1970 Mar;137(1):84–90. doi: 10.1016/0003-9861(70)90413-3. [DOI] [PubMed] [Google Scholar]
  2. Anderson L. E., Fuller R. C. Photosynthesis in Rhodospirillum rubrum. IV. Isolation and characterization of ribulose 1,5-diphosphate carboxylase. J Biol Chem. 1969 Jun 25;244(12):3105–3109. [PubMed] [Google Scholar]
  3. BOARDMAN N. K. Studies on a protochlorophyll-protein complex. I. Purification and molecular-weight determination. Biochim Biophys Acta. 1962 Jul 30;62:63–79. doi: 10.1016/0006-3002(62)90492-4. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Hedrick J. L., Smith A. J. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968 Jul;126(1):155–164. doi: 10.1016/0003-9861(68)90569-9. [DOI] [PubMed] [Google Scholar]
  6. Huffaker R. C., Obendorf R. L., Keller C. J., Kleinkopf G. E. Effects of Light Intensity on Photosynthetic Carboxylative Phase Enzymes and Chlorophyll Synthesis in Greening Leaves of Hordeum vulgare L. Plant Physiol. 1966 Jun;41(6):913–918. doi: 10.1104/pp.41.6.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. KUPKE D. W. Correlation of a soluble leaf protein with chlorophyll accumulation. J Biol Chem. 1962 Oct;237:3287–3291. [PubMed] [Google Scholar]
  8. Kieras F. J., Haselkorn R. Properties of ribulose-1,5-diphosphate carboxylase (carboxydismutase) from Chinese cabbage and photosynthetic microorganisms. Plant Physiol. 1968 Aug;43(8):1264–1270. doi: 10.1104/pp.43.8.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. MacElroy R. D., Johnson E. J., Johnson M. K. Characterization of ribulose diphosphate carboxylase and phosphoribulokinase from Thiobacillus thioparus and Thiobacillus neapolitanus. Arch Biochem Biophys. 1968 Sep 20;127(1):310–316. doi: 10.1016/0003-9861(68)90231-2. [DOI] [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. TROWN P. W. AN IMPROVED METHOD FOR THE ISOLATION OF CARBOXYDISMUTASE. PROBABLE IDENTITY WITH FRACTION I PROTEIN AND THE PROTEIN MOIETY OF PROTOCHLOROPHYLL HOLOCHROME. Biochemistry. 1965 May;4:908–918. doi: 10.1021/bi00881a018. [DOI] [PubMed] [Google Scholar]
  13. WEISSBACH A., HORECKER B. L., HURWITZ J. The enzymatic formation of phosphoglyceric acid from ribulose diphosphate and carbon dioxide. J Biol Chem. 1956 Feb;218(2):795–810. [PubMed] [Google Scholar]

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

RESOURCES