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Photosynthesis Research

, Volume 35, Issue 1, pp 55–66 | Cite as

Redox regulation of enzymatic activity and proteolytic susceptibility of ribulose-1,5-bisphosphate carboxylase/oxygenase fromEuglena gracilis

  • Carlos García-Ferris
  • Joaquín Moreno
Regular Paper

Abstract

The activity of ribulose-1,5-bisphosphate carboxylase/oxygenase fromEuglena gracilis decays steadily when exposed to agents that induce oxidative modification of cysteine residues (Cu2+, benzofuroxan, disulfides, arsenite, oxidized ascorbate). Inactivation takes place with a concomitant loss of cysteine sulfhydryl groups and dimerization of large subunits of the enzyme. 40% activity loss induced by the vicinal thiol-reagent arsenite is caused by modification of a few neighbor residues while the almost complete inactivation achieved with disulfides is due to extensive oxidation leading to formation of mixed disulfides with critical cysteines of the protein. In most cases oxidative inactivation is also accompanied by an increased sensitivity to proteolysis by trypsin, chymotrypsin or proteinase K. Both enzymatic activity and resistance to proteolysis can be restored through treatment with several thiols (cysteamine, cysteine, dithiothreitol and, more slowly, reduced glutathione). Redox effectors which are thought to regulate the chloroplast activity (NADPH, ferredoxin and thioredoxin) do not reactivate the oxidized enzyme. When ribulose-1,5-bisphoshate carboxylase/oxygenase is incubated with cystamine/cysteamine mixtures having different disulfide/thiol ratio (r), inactivation takes place around r=1.5 while proteolytic sensitization occurs under more oxidative conditions (r=4). It is suggested that oxidative modification may happen in vivo under exceptional circumstances, such as senescence, bleaching or different kinds of stress, leading to enzyme inactivation and triggering the selective degradation of the carboxylase that has been repeatedly observed during these processes.

Key words

cysteine oxidation Euglena gracilis protein turnover proteolytic susceptibilization ribulose-1,5-bisphosphate carboxylase/oxygenase 

Abbreviations

CSH

cysteamine

CSSC

cystamine

DTNB

5,5′-dithiobis(2-nitrobenzoic acid)

DTT

dithiothreitol

GSH

reduced glutathione

GSSG

oxidized glutathione

ribulose-P2

ribulose-1,5-bisphosphate

Rubisco

ribulose-1,5-bisphosphate carboxylase/oxygenase

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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Carlos García-Ferris
    • 1
  • Joaquín Moreno
    • 1
  1. 1.Department de Bioquímica i Biologia Molecular, Facultades de CienciasUniversitat de ValènciaBurjassotSpain

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