title: Enzymes as biosensors. 2. Hysteretic response of chloroplastic fructose-1,6-bisphosphatase to fructose 2,6-bisphosphate.
abstract: Oxidized chloroplastic fructose-bisphosphatase is almost totally inactive at pH 7.5, that is under pH conditions that prevail in the chloroplast stroma. When preincubated for different time periods with fructose 2,6-bisphosphate and assayed in the absence of this ligand, it displays an activity which is directly related to the duration of the preincubation phase. This implies that fructose 2,6-bisphosphate induces enzyme conformers that appear in sequence and may be competent for catalytic activity. Upon desorption of fructose 2,6-bisphosphate the enzyme may retain its active conformation for a time period whose duration depends on magnesium concentration. It thus appears that reduction of the enzyme is not an obligatory prerequisite for its activity. Fructose 2,6-bisphosphate behaves as a competitive inhibitor of the reduced, active enzyme, with respect to the real substrate. When assayed with the oxidized enzyme, however, it behaves as an activator. Moreover the apparent steady-state rate that may be measured experimentally depends on both fructose 2,6-bisphosphate concentration and the direction of a concentration change. The reaction velocity experimentally measured is thus a meta-steady-state rate and depends on the initial conditions of the system. The fructose-bisphosphatase system thus displays, with respect to fructose 2,6-bisphosphate, a hysteresis loop and may then sense whether the concentration of that ligand is increased or decreased. A model has been proposed which allows one to explain these results. This model is based on the view that the substrate and fructose 2,6-bisphosphate compete for the same site of the enzyme and that this latter ligand stabilizes a conformation competent for enzyme activity. After the ligand has been chased away, the enzyme retains the active conformation for a while and slowly relapses to the initial inactive conformation. The time-scale of this slow relaxation overlaps that of the steady state of product appearance and this generates meta-steady-state kinetics, which is dependent on the initial state and therefore on the history of the system.
authors.affiliation: Centre de Biochimie et de Biologie Moléculaire du CNRS, Marseille, France.
authors.forename: J M
authors.initials: JM
authors.lastname: Soulie
authors.forename: M
authors.initials: M
authors.lastname: Riviere
authors.forename: J
authors.initials: J
authors.lastname: Ricard
mesh_terms: D001667:Binding, Competitive; D002736:Chloroplasts / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D006597:Fructose-Bisphosphatase / Q000378:metabolism*; D005635:Fructosediphosphates / Q000378:metabolism* / Q000494:pharmacology; D006598:Hexosediphosphates / Q000378:metabolism*; D008024:Ligands; D008956:Models, Chemical; D010084:Oxidation-Reduction; D011485:Protein Binding; D011487:Protein Conformation; D013816:Thermodynamics; D013997:Time Factors
chemical_list: D005635:Fructosediphosphates; D006598:Hexosediphosphates; D008024:Ligands; C027652:fructose 2,6-diphosphate; D006597:Fructose-Bisphosphatase