Activation of cyanobacterial RuBP-carboxylase/oxygenase is facilitated by inorganic phosphate via two independent mechanisms

Yehouda Marcus, Michael Gurevitz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Orthophosphate (P(i)) modulates the activity and activation of ribulose 1,5-bis-phosphate carboxylase/oxygenase (RuBisCO) via a mechanism that is still controversial. Whereas its effects on the higher plant enzyme have been described, little is known about P(i) regulation of the structurally similar, yet kinetically different cyanobacterial enzyme. We found that RuBisCO of Synechocystis PCC6803 was affected by P(i) in a paradoxical fashion. On the one hand, P(i) inhibited catalysis by competing with the substrate RuBP, and on the other hand it stimulated enzyme activation in a dual manner manifested by multiphasic kinetics, which differed from the effect on activation of the higher plant enzyme. P(i) concentrations > 5 mM promoted the carbamylation of the cyanobacterial enzyme and the binding of Mg2+ to the carbanion at suboptimal concentrations of CO2 and Mg2+. Surprisingly, P(i) also increased the activation level of the carbamylated enzyme via another putative site of interaction. In contrast with the higher plant RuBisCO, RuBP did not inhibit the stimulatory effect of phosphate on activation of the cyanobacterial enzyme, suggesting a P(i) effect through a site other than the sugar binding site. The dual effect on activation could be distinguished by the phosphate analogue vanadate, which inhibited only the stimulation achieved at high phosphate concentrations. The elevation of RuBisCO activation at suboptimal levels of CO2 and high concentrations of RuBP suggests that in cyanobacteria P(i) may have a role analogous to that of RuBisCO activase in higher plants.

Original languageEnglish
Pages (from-to)5995-6003
Number of pages9
JournalEuropean Journal of Biochemistry
Volume267
Issue number19
DOIs
StatePublished - 2000

Keywords

  • Cyanobacteria
  • Phosphate
  • RuBisCO activation
  • Vanadate

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