Kinetic model for the action of the inorganic pyrophosphatase from Streptococcus faecalis.
Kinetic studies of the less active form of Streptococcus faecalis inorganic pyrophosphatase (EC 3.6.1.1), together with computational analysis, indicated that cooperativity in ligand binding contributes in a significant way to the behavior of this enzyme. The simplest model applicable to our data was a Monod-Wyman-Changeux-type, allosteric model, in which the enzyme is proposed to exist in two states, referred to as R and T states, respectively. In the absence of ligands, 94% of the enzyme was in the T state. MgPPi2- was the only substrate for the enzyme in the R form. This substrate was bound equally well by both enzyme forms, but it was hydrolyzed 5 times more efficiently by the R form than it was by the T form. Mg2PPi was bound exclusively to the T state of the enzyme, and it was hydrolyzed 25% as rapidly as MgPPi2- by the T form. Mg2PPi inhibited the hydrolysis of the more efficient substrate, MgPPi2-, by competing with MgPPi2- for the enzyme in the T form and by shifting the R----T equilibrium in favor of the T form. Mg2+ stabilized the R state, thus activating the hydrolysis of MgPPi2- and inhibiting that of Mg2PPi.
D013293:Enterococcus faecalis / Q000201:enzymology*; D043564:Inorganic Pyrophosphatase; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D008954:Models, Biological; D011485:Protein Binding; D011755:Pyrophosphatases / Q000378:metabolism*
D046911:Macromolecular Substances; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase