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Simple determination of the carbon dioxide to oxygen ratio specificity of ribulose 1 5 bisphosphate carboxylase oxygenase by the specific radioactivity of carbon 14 glycerate 3 phosphate



Simple determination of the carbon dioxide to oxygen ratio specificity of ribulose 1 5 bisphosphate carboxylase oxygenase by the specific radioactivity of carbon 14 glycerate 3 phosphate



Plant Physiology 98(2): 764-768



A new method is presented for measurement of the CO2/O2 specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The [14C]3-phosphoglycerate (PGA) from the Rubisco carboxylase reaction and its dilution by the Rubisco oxygenase reaction was monitored by directly measuring the specific radioactivity of PGA. 14CO2 fixation with Rubisco occurred under two reaction conditions: carboxylase with oxygenase with 40 micromolar CO2 in O2-saturated water and carboxylase only with 160 micromolar CO2 under N2. Detection of the specific radioactivity used the amount of PGA as obtained from the peak area, which was determined by pulsed amperometry following separation by high-performance anion exchange chromatography and the radioactive counts of the [14C]PGA in the same peak. The specificity factor of Rubisco from spinach (Spinacia oleracea L.) (93 .+-. 4), from the green alga Chlamydomonas reinhardtii (66 .+-. 1), and from the photosynthetic bacterium Rhodospirillum rubrum (13) were comparable with the published values measured by different methods.

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