Kinetic and thermodynamic study of the tetramerization equilibrium of phosphorylase b.
The association equilibrium of phosphorylase b, induced by AMP and in the presence of Mg2+, has been shown to be a reversible process that follows second order and first order reversible rate laws in the direction of tetramerization and dimerization respectively, this fact being independent of temperature and of enzyme and AMP concentrations. Moreover, rate and equilibrium constants have been evaluated and their dependence on temperature and AMP concentration studied in this work. An important role that the existence of two classes of AMP binding sites per enzymatic subunit plays in the aggregation properties of the enzyme has also been emphasized. In the presence of 0.1 and 1 mM AMP (binding to the high affinity site), the values of the change in enthalpy, activation energy of dimerization and activation energy of tetramerization are: -36 kcal/mol, +36 kcal/mol, and 0 kcal/mol respectively. Binding of AMP to the low affinity site (10 mM AMP) yields significant changes in the self-association equilibrium, since the preceding parameters reach the following values: -18, +32, and +14 kcal/mol.
D000249:Adenosine Monophosphate; D000818:Animals; D001704:Biopolymers; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D007700:Kinetics; D008274:Magnesium; D010763:Phosphorylase b; D006005:Phosphorylases; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D013696:Temperature; D013816:Thermodynamics
D001704:Biopolymers; D000249:Adenosine Monophosphate; D010763:Phosphorylase b; D006005:Phosphorylases; D008274:Magnesium