title: Thioredoxin activation of phosphoribulokinase in a bi-enzyme complex from Chlamydomonas reinhardtii chloroplasts.
abstract: The activation of oxidized phosphoribulokinase either "free" or as part of a bi-enzyme complex by reduced thioredoxins during the enzyme reaction was studied. In the presence of reduced thioredoxin, the product of the reaction catalyzed by phosphoribulokinase within the bi-enzyme complex does not appear in a linear fashion. It follows a mono-exponential pattern that suggests a slow dissociation process of the bi-enzyme complex in the assay cuvette. A plot of the steady state of product appearance against thioredoxin concentration gave a sigmoid curve. On the basis of our experimental results, we propose a minimum model of the activation of phosphoribulokinase by reduced thioredoxin. Reduced thioredoxin may act on the phosphoribulokinase, either within the complex or in the dissociated metastable form. However, the time required to activate the enzyme as part of the complex is shorter (about 20 s) than that required to activate the dissociated form (about 10 min). This might be of physiological relevance, and we discuss the role of the interactions between phosphoribulokinase and glyceraldehyde-3-phosphate dehydrogenase in the regulation of the Calvin cycle.
authors.affiliation: Institut Jacques MONOD (UMR 7592), CNRS-Universités Paris VI-VII, 2 Place Jussieu, 75251 Paris Cedex 05, France.
authors.forename: L
authors.initials: L
authors.lastname: Avilan
authors.forename: S
authors.initials: S
authors.lastname: Lebreton
authors.forename: B
authors.initials: B
authors.lastname: Gontero
mesh_terms: D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000201:enzymology*; D002736:Chloroplasts / Q000201:enzymology*; D004789:Enzyme Activation; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D007700:Kinetics; D008956:Models, Chemical; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*
chemical_list: D013879:Thioredoxins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D017853:Phosphotransferases (Alcohol Group Acceptor); C022390:phosphoribulokinase