Electron spin resonance study of human alpha 1-acid glycoprotein interaction with a spin labelled steroid.
The interaction of human alpha 1-acid glycoprotein (AAG) with a corticosteroid was studied using nitroxide labeled deoxycorticosterone and electron spin resonance (ESR) spectroscopy. The ESR spectra of the spin labeled steroid in the presence of AAG could be used to characterize the ligand-protein interaction at equilibrium without the need of a separation between bound and free species. An association constant Ka of 6.10(5) M-1 at 20 degrees C and a binding capacity of one site per mole protein were found. ESR spectra recorded at equilibrium at various temperatures allowed the calculation of enthalpy and entropy variations for the steroid-protein interaction; these thermodynamic parameters exhibited a rapid change above 45 degrees C which may be related to a protein conformational modification above this temperature, as detected by circular dichroism study. The ESR spectra width could be used to define a polar character for the spin label environment in the steroid binding site of AAG and to calculate an apparent rotational correlation time of 2.8 x 10(-8) sec for the steroid-protein complex in aqueous solution at 20 degrees C. It can be concluded that spin labeling and ESR methodology is of value in the study of steroid-protein interactions of biological significance above all because it can provide direct physico-chemical information concerning the local environment of the ligand in its binding site at equilibrium.
D001665:Binding Sites / Q000187:drug effects; D002942:Circular Dichroism; D003345:Corticosterone / Q000031:analogs & derivatives; D003900:Desoxycorticosterone / Q000031:analogs & derivatives; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D006801:Humans; D006854:Hydrocortisone; D009961:Orosomucoid; D012399:Rotation; D013113:Spin Labels; D013816:Thermodynamics
D009961:Orosomucoid; D013113:Spin Labels; D003900:Desoxycorticosterone; C026367:corticosterone nitroxide; C026366:deoxycorticosterone nitroxide; D003345:Corticosterone; D006854:Hydrocortisone