Thermodynamics of nucleotides binding to phosphorylase b.
The effects of several chemical modifications in the AMP molecule on its interaction with phosphorylase b are examined by microcalorimetry, equilibrium dialysis, light scattering and ultracentrifuge experiments. In this work we report the results obtained for eight AMP analogues corresponding to different substituents in the puric base or in the ribose, or to different positions of the phosphate. The thermodynamic properties of the interaction between the phosphorylase b and the above mentioned nucleotides are also reported. The following conclusions were obtained: a) Except for IMP and 2'dIMP all the nucleotides studied clearly show two types of binding sites in the enzyme. b) The interaction of the nucleotide with its weaker affinity binding site is highly dependent upon chemical alterations in the puric base. c) Both the amino group in C(6) and the N(1) of the adenine in the AMP seem to play an important role in the conformational transitions induced by the nucleotide on the enzyme. d) The tetramerization of phosphorylase b in the presence of 10(-2) M AMP and in the conditions of the ultracentrifuge experiments is drastically affected by modifications in the ribose-phosphate part of the AMP molecule.
D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D007700:Kinetics; D009132:Muscles / Q000201:enzymology; D010763:Phosphorylase b; D006005:Phosphorylases; D011485:Protein Binding; D011817:Rabbits; D012265:Ribonucleotides; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D012265:Ribonucleotides; D010763:Phosphorylase b; D006005:Phosphorylases