Assay of nucleotides and other phosphate-containing compounds in isolated chloroplasts by ion exchange chromatography
✍ Scribed by Hans W. Heldt; Archie R. Portis; Ross McC.Lilley; Annemarie Mosbach; Chong Ja Chon
- Publisher
- Elsevier Science
- Year
- 1980
- Tongue
- English
- Weight
- 683 KB
- Volume
- 101
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
This report describes a method for a sensitive assay of phosphate-containing metabolites in chloroplasts. The first step was to preincubate the chloroplasts with inorganic [3ZP]phosphate in order to label the stromal phosphate pool. On illumination, the intermediates of the reductive CO, fixation cycle, and also ADP and ATP, were radioactively labeled. After a metabolic stop by filtering centrifugation into perchloric acid, the resulting extract was chromatographed on a small ion exchange column with a formic acid-formate gradient. The separated substances which appeared in the eluate were detected by continuous monitoring of 32P radioactivity. The identity of the peaks in the chromatogram and the reproducibility of the assay procedure were investigated.