Is leaf ADP-glucose pyrophosphorylase an allosteric enzyme?
Barley leaf ADP-glucose pyrophosphorylase (AGPase), a key enzyme of starch synthesis in the chloroplast stroma, was analysed, in both directions of the reaction, with respect to details of its regulation by 3-phosphoglycerate (PGA) and inorganic phosphate (Pi) which serve as activator and inhibitor, respectively. AGPase was found to catalyse a close-to-equilibrium reaction, with the K(eq) value of approximately 0.5, i.e. slightly favouring the pyrophosphorolytic direction. When the enzyme was analysed by substrate kinetics, PGA acted either as a linear (hyperbolic response) 'non-competitive' activator (forward reaction) or a linear near-'competitive' activator (reverse reaction). When the activation and inhibition patterns with PGA and Pi, respectively, were studied in detail by Dixon plots, the response curves to effectors also followed hyperbolic kinetics, with the experimentally determined K(a) and K(i) values on the order of micromolar. The results suggest that the regulation of AGPase proceeds via a non-cooperative mechanism, where neither of the effectors, when considered separately, induces any allosteric response. The evidence, discussed in terms of an overall kinetic mechanism/regulation of leaf AGPase, prompts caution in classifying the protein as an 'allosteric enzyme'.
Kleczkowski
L A
LA
Umeå Plant Science Centre, Department of Plant Physiology, Umeâ University, 901 87, Umeå, Sweden. lesek.kleczkowsi@plantphys.umu.se
D000494:Allosteric Regulation; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051599:Glucose-1-Phosphate Adenylyltransferase; D005988:Glyceric Acids / Q000494:pharmacology; D001467:Hordeum; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000737:chemistry*; D010710:Phosphates / Q000494:pharmacology; D018515:Plant Leaves / Q000201:enzymology
D004791:Enzyme Inhibitors; D005988:Glyceric Acids; D010710:Phosphates; C005156:3-phosphoglycerate; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase