Ligand-induced asymmetry as observed through fluorophore rotations and free energy couplings: application to neurophysin.
Changes that occur in subunit neurophysin structure upon ligand binding were explored by two methods. First, the thermal coefficient of the viscosity around the subunit tyrosine was monitored, which yields information on the environmental flexibility and free rotational space of the fluorophore. Initially, it was determined that the environmental flexibility and the free space around each subunit tyrosine are unperturbed upon dimerization. Binding of the tripeptide analogue of oxytocin causes the once homologous environments of the subunit tyrosines to become drastically different such that one moves onto a closely packed environment whereas the other moves into a region of larger free space. Even though the subunits as seen by each tyrosine are very different, the specific binding sites as seen by the ligands are similar. It was also found that ligand binding is stabilized by ring stacking and that energy transfer occurs between the tyrosine of the ligand and the neurophysin subunit tyrosine. Second, changes in subunit structure upon ligation were also followed by the determination of the order of free energy coupling between ligand binding and oligomerization, which tells how each ligand affects the subunit affinity. Since the binding of ligand is cooperative and induces dimer formation, there is second-order coupling between ligand binding and dimerization and the binding of the second ligand is responsible for the increase in subunit affinity.
D000818:Animals; D002417:Cattle; D007700:Kinetics; D008024:Ligands; D046911:Macromolecular Substances; D009481:Neurophysins; D009842:Oligopeptides; D011485:Protein Binding; D011487:Protein Conformation; D013816:Thermodynamics; D014443:Tyrosine; D014783:Viscosity
D008024:Ligands; D046911:Macromolecular Substances; D009481:Neurophysins; D009842:Oligopeptides; D014443:Tyrosine