Abstract
Cell-free extracts of Prochloron didemni were assayed for ribulose 1,5-bisphosphate (RuBP) carboxylase (EC 4.1.1.39) and phosphoribulokinase (EC 2.7.1.19), two key enzymes in the reductive pentose-phosphate cycle. In an RuBP-dependent reaction, the production of two molecules of 3-phosphoglycerate per molecule of CO2 fixed was shown. Phosphoribulokinase activity was demonstrated by the production of ADP from ribulose 5-phosphate (Ru5P) and ATP and by measurement of ATP-, Ru5P-dependent 14CO2 fixation in the presence of excess spinach RuBP carboxylase. When Prochloron RuBP carboxylase was purified from cell-free extracts by isopycnic centrifugation in reoriented linear 0.2 to 0.8 M sucrose gradients, the enzyme sedimented to a position which corresponded to that for the 520,000-dalton spinach enzyme. After polyacrylamide gel electrophoresis (PAGE) of Prochloron enzyme, a major band of enzyme activity corresponded to that for the spinach enzyme. Considerably more additional carboxylase activity was found in a less mobile species than was the case for spinach RuBP carboxylase. Sodium dodecyl sulfate-PAGE of the Prochloron enzyme indicates that it is composed of both large (molecular weight, MW=57,500) and small (MW=18,800) subunits.
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Abbreviations
- PAGE:
-
polyacrylamide gel electrophoresis
- PGA:
-
3-phospho D-glycerate
- RuBP:
-
D-ribulose 1,5-bisphosphate
- Ru5P:
-
D-ribulose 5-phosphate
- SDS:
-
sodium dodecylsulfate
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Berhow, M.A., McFadden, B.A. Ribulose 1,5-bisphosphate carboxylase and phosphoribulokinase in Prochloron . Planta 158, 281–287 (1983). https://doi.org/10.1007/BF00397328
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DOI: https://doi.org/10.1007/BF00397328