Abstract
An inverse relationship between the concentration of ribose 5-phosphate and apparent ribulose bisphosphate carboxylase activity was observed. The Lilley-Walker assay spectrophotometric assay, in which the 3-phosphoglyceric acid-dependent oxidation of reduced pyridine nucleotide is measured, is shown to be highly sensitive to inhibition by heavy metals. Analysis of the purity of reagents showed that ribose 5-phosphate is often contaminated with lead in sufficient quantity to impair the assay. This noncompetitive inhibition by ribose 5-phosphate is independent of the competitive inhibition of this substrate as an ATP sink as described by Slabas and Walker. A method for checking reagent purity and removing heavy metal contaminants is described.
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- Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Larsson-Raźnikiewicz M., Arvidsson L. Inhibition of phosphoglycerate kinase by products and product homologues. Eur J Biochem. 1971 Oct 26;22(4):506–512. doi: 10.1111/j.1432-1033.1971.tb01570.x. [DOI] [PubMed] [Google Scholar]
- Lilley R. M., Walker D. A. An improved spectrophotometric assay for ribulosebisphosphate carboxylase. Biochim Biophys Acta. 1974 Jul 17;358(1):226–229. doi: 10.1016/0005-2744(74)90274-5. [DOI] [PubMed] [Google Scholar]
- Slabas A. R., Walker D. A. Localization of inhibition by adenosine diphosphate of phosphoglycerate-dependent oxygen evolution in a reconstituted chloroplast system. Biochem J. 1976 Jan 15;154(1):185–192. doi: 10.1042/bj1540185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slabas A. R., Walker D. A. Transient inhibition by ribose 5-phosphate of photosynthetic O2 evolution in a reconstituted chloroplast system. Biochim Biophys Acta. 1976 Apr 9;430(1):154–164. doi: 10.1016/0005-2728(76)90231-0. [DOI] [PubMed] [Google Scholar]