Abstract
A procedure is described to prepare uniformly labelled D-[14C]ribulose 1,5-bisphosphate enzymically from uniformly labelled D-[14C]glucose through the coupled reactions catalysed by hexokinase (EC 2.7.1.1), glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44) and 5-phosphoribulokinase (EC 2.7.1.19). All reagents utilized in the method are commercially available. The procedure is a reliable preparative-scale method for synthesizing the dibarium salt of D-[14C]ribulose 1,5-biphosphate with a specific radioactivity up to 7 mCi/mmol and a purity near 90%. The final product was free of other 14C-labelled sugars, sugar phosphate esters, Pi and nucleotides.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andrews T. J., Lorimer G. H., Tolbert N. E. Ribulose diphosphate oxygenase. I. Synthesis of phosphoglycolate by fraction-1 protein of leaves. Biochemistry. 1973 Jan 2;12(1):11–18. doi: 10.1021/bi00725a003. [DOI] [PubMed] [Google Scholar]
- Christen P. Paracatalytic enzyme modification by oxidation of enzyme-substrate carbanion intermediates. Methods Enzymol. 1977;46:48–54. doi: 10.1016/s0076-6879(77)46009-9. [DOI] [PubMed] [Google Scholar]
- HANES C. S., ISHERWOOD F. A. Separation of the phosphoric esters on the filter paper chromatogram. Nature. 1949 Dec 31;164(4183):1107-12, illust. doi: 10.1038/1641107a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Kuehn G. D. Uridine diphosphoglucose pyrophosphorylase activity and differentiation in the cellular slime mold Physarum polycephaluno. J Bacteriol. 1974 Dec;120(3):1151–1157. doi: 10.1128/jb.120.3.1151-1157.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- TYSZKIEWICZ E. An improved solvent system for the paper chromatography of phosphate esters. Anal Biochem. 1962 Feb;3:164–166. doi: 10.1016/0003-2697(62)90107-0. [DOI] [PubMed] [Google Scholar]
- Wishnick M., Lane M. D. Inhibition of ribulose diphosphate carboxylase by cyanide. Inactive ternary complex of enzyme, ribulose diphosphate, and cyanide. J Biol Chem. 1969 Jan 10;244(1):55–59. [PubMed] [Google Scholar]