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Title: Simple determination of the CO sub 2 /O sub 2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of ( sup 14 C) glycerate 3-phosphate

Abstract

A new method is presented for measurement of the CO{sub 2}/O{sub 2} specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The ({sup 14}C)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. {sup 14}CO{sub 2} fixation with Rubisco occurred under two reaction conditions: carboxylase with oxygenase with 40 micromolar CO{sub 2} in O{sub 2}-saturated water and carboxylase only with 160 micromolar CO{sub 2} under N{sub 2}. 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 ({sup 14}C)PGA in the same peak. The specificity factor of Rubisco from spinach (Spinacia oleracea L.) (93 {plus minus} 4), from the green alga Chlamydomonas reinhardtii (66 {plus minus} 1), and from the photosynthetic bacterium Rhodospirillum rubrum (13) were comparable with the published values measured by different methods.

Authors:
; ; ;  [1]
  1. Univ. of Arizona, Tucson (United States)
Publication Date:
OSTI Identifier:
7236274
Resource Type:
Journal Article
Journal Name:
Plant Physiology; (United States)
Additional Journal Information:
Journal Volume: 98:2; Journal ID: ISSN 0032-0889
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; RIBULOSE DIPHOSPHATE CARBOXYLASE; SPECIFICITY; BIOLOGICAL PATHWAYS; CARBON 14 COMPOUNDS; CARBON DIOXIDE; CARBOXYLATION; CHLAMYDOMONAS; OXYGEN; PHOTOSYNTHETIC BACTERIA; RADIOACTIVITY; SPINACH; TRACER TECHNIQUES; ALGAE; CARBON COMPOUNDS; CARBON OXIDES; CARBON-CARBON LYASES; CARBOXY-LYASES; CHALCOGENIDES; CHEMICAL REACTIONS; CHLOROPHYCOTA; ELEMENTS; ENZYMES; FOOD; ISOTOPE APPLICATIONS; LABELLED COMPOUNDS; LYASES; MAGNOLIOPHYTA; MAGNOLIOPSIDA; MICROORGANISMS; NONMETALS; ORGANIC COMPOUNDS; OXIDES; OXYGEN COMPOUNDS; PLANTS; PROTEINS; UNICELLULAR ALGAE; VEGETABLES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Zhu, Genhai, Jensen, R G, Hallick, R B, and Wildner, G F. Simple determination of the CO sub 2 /O sub 2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of ( sup 14 C) glycerate 3-phosphate. United States: N. p., 1992. Web.
Zhu, Genhai, Jensen, R G, Hallick, R B, & Wildner, G F. Simple determination of the CO sub 2 /O sub 2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of ( sup 14 C) glycerate 3-phosphate. United States.
Zhu, Genhai, Jensen, R G, Hallick, R B, and Wildner, G F. Sat . "Simple determination of the CO sub 2 /O sub 2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of ( sup 14 C) glycerate 3-phosphate". United States.
@article{osti_7236274,
title = {Simple determination of the CO sub 2 /O sub 2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of ( sup 14 C) glycerate 3-phosphate},
author = {Zhu, Genhai and Jensen, R G and Hallick, R B and Wildner, G F},
abstractNote = {A new method is presented for measurement of the CO{sub 2}/O{sub 2} specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The ({sup 14}C)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. {sup 14}CO{sub 2} fixation with Rubisco occurred under two reaction conditions: carboxylase with oxygenase with 40 micromolar CO{sub 2} in O{sub 2}-saturated water and carboxylase only with 160 micromolar CO{sub 2} under N{sub 2}. 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 ({sup 14}C)PGA in the same peak. The specificity factor of Rubisco from spinach (Spinacia oleracea L.) (93 {plus minus} 4), from the green alga Chlamydomonas reinhardtii (66 {plus minus} 1), and from the photosynthetic bacterium Rhodospirillum rubrum (13) were comparable with the published values measured by different methods.},
doi = {},
url = {https://www.osti.gov/biblio/7236274}, journal = {Plant Physiology; (United States)},
issn = {0032-0889},
number = ,
volume = 98:2,
place = {United States},
year = {1992},
month = {2}
}